Team:UT Dallas/week5

From 2013.igem.org

8/1

<a name="h.gdt42dr1sitx"></a>Miniprep

<a name="h.k16mhl32quoi"></a>i5-16-1 [Com B]                7.4

<a name="h.ykgo7uctfizd"></a>i5-16-2 [Com B]                5.4

<a name="h.b5syoehwe5sd"></a>i5-16-3[Com B]                13.7

<a name="h.mfxia9twmnj"></a>i5-16-4 [Scr]                no growth

<a name="h.601z1zffypq"></a>i5-16-5 [Scr]                10.7

<a name="h.ycdqtoz6j1jh"></a>i5-16-6[Scr]                7.8

<a name="h.ajykp6mwqppf"></a>i5-16-7[pScr]                8.4

<a name="h.ne55wp3gkau3"></a>i5-16-8[pScr]                9.1

<a name="h.e13d54dyzft7"></a>i5-16-9[pScr]                10.6

<a name="h.txo346njagbb"></a>i5-16-10 [Cra]                19.9

<a name="h.c3473p5uadpe"></a>i5-16-11[Cra]                5.2

<a name="h.wbmb3g1s7ugf"></a>i5-16-12 [Cra]                6.4

<a name="h.kv0tfbo2jv8i"></a>i5-16-13[pFru]                15.3

<a name="h.j3t9j0u7xkkq"></a>i5-16-14[pFru]                5.4

<a name="h.ilxeqahhep0j"></a>i5-16-15[pFru]                5.3

<a name="h.iwp9ty6hlxs1"></a>i5-16-16[Com C]        2.4

<a name="h.n4f67hryzt17"></a>i5-16-17[Com C]        7.7

<a name="h.l8oo77q6fsr"></a>i5-16-18[Com C]        5.8

<a name="h.df02fsoc2gkk"></a>

<a name="h.s0shikkagflk"></a>Concentrations too low

<a name="h.h0j00xjiuue9"></a>Redo digestion and ligation (Plan B)

<a name="h.7k5mc2xcbp6i"></a>

<a name="h.1h2u7hs3ix23"></a>Digestion

<a name="h.g3z76xurwd39"></a>

<a href="#" name="fdd2c2cc1a30f21ef2d5a146b718253990fe43a8"></a><a href="#" name="13"></a><tbody></tbody>

<a name="h.nw7x4tp0kj95"></a>

<a name="h.b77riogep97k"></a>i5-11-5 (100.9 ng/ul) [pLac-RBS-GFP]

<a name="h.7x23o2q63rld"></a>DNA

<a name="h.xs95dkb0qv3g"></a>20

<a name="h.qmbaclu4and5"></a>NEBuffer

<a name="h.i4e84vgkrcxx"></a>5

<a name="h.9fr4omfw11ah"></a>EcoR

<a name="h.xp4lm573njg7"></a>1

<a name="h.fnra7wygpvwz"></a>PstI

<a name="h.k3078dha6squ"></a>1

<a name="h.uzdcyicnhuw5"></a>Water

<a name="h.peiv6cad45bh"></a>23

<a name="h.wla559qqazui"></a>Total

<a name="h.snx6esd4tjh1"></a>50 → i5-17-1

<a name="h.hxsbfqaq1flk"></a>

<a name="h.ak8khbywm050"></a>Digestion

<a name="h.qpk8acfchyb7"></a>

<a href="#" name="7150480a806c7acfe65074c44a5a56b72df44278"></a><a href="#" name="14"></a><tbody></tbody>

<a name="h.plaqos35rnbu"></a>

<a name="h.oduap9peuhwz"></a>i4-45-3

<a name="h.28slfgkpyrku"></a>i4-45-2

<a name="h.qm7ywscmvd1o"></a>i5-11-5 [pLac-RBS-GFP]

<a name="h.2y6nakdykx4n"></a>DNA

<a name="h.nluimw5ad5ii"></a>20

<a name="h.euarp7r4qqr4"></a>20

<a name="h.u7pjsz33yp8u"></a>20

<a name="h.fnxqh2ibdalu"></a>NEBuffer

<a name="h.kme0h754mt8u"></a>5

<a name="h.pychv9g0t5aj"></a>5

<a name="h.90qha3jcqgai"></a>5

<a name="h.xp8k4bpokxpe"></a>EcoR

<a name="h.simmuu8727lv"></a>1

<a name="h.d4s682l63gvk"></a>1

<a name="h.g0zvv2bkjcot"></a>1

<a name="h.lctki6qvqypt"></a>PstI

<a name="h.mvrdl2k9tfm"></a>1

<a name="h.e1qgxndsr3jw"></a>1

<a name="h.pcbwtt3jlera"></a>1

<a name="h.kn30n1rxa7pm"></a>Water

<a name="h.hfipwz68ut8i"></a>23

<a name="h.9a1getpgeol4"></a>23

<a name="h.xfxc3bw7xdpb"></a>23

<a name="h.7ubx7vr86r15"></a>Total

<a name="h.2fqkbpkycpxr"></a>50

<a name="h.cd74i22n9ixv"></a>50

<a name="h.2thgvsmqsnes"></a>50

<a name="h.rnkzjpj51cq2"></a>

<a name="h.462qh4ufcpyq"></a>Nanodrop i4-45-2        18.4

<a name="h.qymzp02bzgjz"></a>        i4-45-3                28.8

<a name="h.58tertazj0wl"></a>

<a name="h.8yjdiim94mo2"></a>Ligation

<a name="h.odd5alle0l7u"></a>Mastermix: Vector: 16         Buffer: 16

<a name="h.9ljnk7muefc"></a>

<a name="h.750mg81plwxp"></a>Vector: i4-45-3

<a name="h.qky6dflkxf5u"></a>

<a href="#" name="d7efde7569928a4e7ee8fc236d3137185b88940d"></a><a href="#" name="15"></a><tbody></tbody>

<a name="h.ko16dp6g3zlu"></a>Insert

<a name="h.g9oyoz6rhqne"></a>i5-13-9

<a name="h.ko16dp6g3zlu"></a>i5-13-9

<a name="h.ko16dp6g3zlu"></a>i5-13-10

<a name="h.g9oyoz6rhqne"></a>i5-13-11

<a name="h.hgf7vl6itpa4"></a>i5-13-12

<a name="h.ke9rrrsnsyde"></a>i5-13-13

<a name="h.1w7eymqeeklq"></a>i5-15-1

<a name="h.ldquz8vbyhe6"></a>

<a name="h.htbyr1ll042h"></a>3:1

<a name="h.qlmj2ypxhaup"></a>0:1

<a name="h.xb0vn1n1rtbt"></a>3:1

<a name="h.ws93oogubgzu"></a>3:1

<a name="h.n5puy2qf1f8"></a>3:1

<a name="h.oikpvcs0iovu"></a>3:1

<a name="h.1hj1ujr3oyqe"></a>3:1

<a name="h.gnq4j8mok285"></a>Insert

<a name="h.p7qkdz74u435"></a>7.5

<a name="h.do5uf7x3xfce"></a>0

<a name="h.gjadgexwrr8k"></a>6

<a name="h.qv4hhw4bo7xq"></a>1

<a name="h.dj2b0xeslc0r"></a>5

<a name="h.wxppe7l5ijoc"></a>1.5

<a name="h.4rr06z9x2bi2"></a>1

<a name="h.6nrozzpemgsl"></a>Vector

<a name="h.294k89lar7ku"></a>2

<a name="h.t20x9jisxas4"></a>2

<a name="h.ihz5jvqdk19p"></a>2

<a name="h.mlguz32wkiza"></a>2

<a name="h.thhdugjt5v1a"></a>2

<a name="h.6iohv0przw1"></a>2

<a name="h.t36qi49njmay"></a>2

<a name="h.ix5ibc2nn2bl"></a>Buffer

<a name="h.rbbw0rx8m512"></a>2

<a name="h.bg1sshpu7d3p"></a>2

<a name="h.dx7eibvymd2p"></a>2

<a name="h.ssu9fv79la3s"></a>2

<a name="h.nmchd34dvlrd"></a>2

<a name="h.o91ingua0t1m"></a>2

<a name="h.67y7n43zvoxa"></a>2

<a name="h.l9lmdir3u5kt"></a>Ligase

<a name="h.y303ooloe8eo"></a>1

<a name="h.f7r14f39zeo9"></a>1

<a name="h.3guxuh1ts81v"></a>1

<a name="h.fco4l7y7m9i0"></a>1

<a name="h.102mclq61u6e"></a>1

<a name="h.wlk08oxitdt"></a>1

<a name="h.f76a5efgbwy0"></a>1

<a name="h.hkl9t5ohw35f"></a>water

<a name="h.unws8s6zidez"></a>7.5

<a name="h.lot17dn25adf"></a>15

<a name="h.pjp8seiw7ctd"></a>9

<a name="h.2njbhf4869si"></a>14

<a name="h.azmz7p4j1n6j"></a>10

<a name="h.i79hrut5w68k"></a>13.5

<a name="h.kuwi6l8k42ow"></a>14

<a name="h.j2utgt1mtyc9"></a>Total

<a name="h.w0wf66iqe0tz"></a>20

<a name="h.7zdyt1ohkrtq"></a>20

<a name="h.l8d72odxorcw"></a>20

<a name="h.i3y4oas8rm7p"></a>20

<a name="h.jrhfboomo7ic"></a>20

<a name="h.7tkqll8rjzny"></a>20

<a name="h.7vhzegboowg7"></a>20

<a name="h.u3kbvtk7cxk"></a>1                2                3                4                5                6                7

<a name="h.mt6betv3gsga"></a>

<a name="h.vdlf9hfxs3za"></a>8/1

<a name="h.3kl77r7ego4"></a>Transformation

<a name="h.qnxw5fcp5bdl"></a>+ control → i5-10-4

<a name="h.rxixlc9jyz7j"></a>

<a name="h.d3hn90r6uogh"></a>1 → 10 colonies → picked 2 → i5-19-1

<a name="h.tky8lnz5box"></a>                                i5-19-2

<a name="h.v9a0276oa2c3"></a>2 →0 colonies

<a name="h.8rg3vllobj6l"></a>3 →0 colonies

<a name="h.terx6m36at4p"></a>4 →20 colonies → picked 2 → i5-19-3

<a name="h.ax82ddgvej52"></a>                                i5-19-4

<a name="h.kl8f3p7w9a94"></a>5 →50 + colonies → picked 2 → i5-19-5

<a name="h.rly6ll3758vh"></a>                                i5-19-6

<a name="h.t6yhc2bwlkmr"></a>6 →0 colonies (ruined plate)

<a name="h.txw92pj0umww"></a>7 →25  colonies → picked 2 → i5-19-7

<a name="h.2pg34ohcmptq"></a>                                i5-19-8

<a name="h.t1r4u7513way"></a>

<a name="h.1yh2ap6r5dkj"></a>8/2

<a name="h.cbb9vyy27u50"></a>Transformation

<a name="h.tkzkibvuxed7"></a>+ control → i5-10-4

<a name="h.ygzhfj4d971e"></a>1 → 6 colonies → picked 2 → i5-19-9

<a name="h.jkleuycfz7gw"></a>                                i5-19-10

<a name="h.92goxt1940i6"></a>3 → 20 colonies → picked 2 → i5-19-11

<a name="h.n4keacizzoq7"></a>                                i5-19-12

<a name="h.1657iecoccwd"></a>6→ 8 colonies → picked 2 → i5-19-13

<a name="h.o4r84ljldi8j"></a>                                i5-19-14

<a name="h.i28qtsm3t7e"></a>

<a name="h.hupfbsd3ivci"></a>8/3

<a name="h.h7xzg2fhesfu"></a>Miniprep

<a name="h.785x68lq4t23"></a>

<a href="#" name="5da75b8ad68cb98f581dda94edaf46683d1aed71"></a><a href="#" name="16"></a><tbody></tbody>

<a name="h.5324zw47di0h"></a>

<a name="h.i1tf65pvidmt"></a>Concentrations ng/ul

<a name="h.v7qvzht8rovt"></a>i5-19-1 [Com B]

<a name="h.oycn5t5zwqw"></a>90.3

<a name="h.yg0vhvy9xbin"></a>i5-19-2 [Com B]

<a name="h.6y6ofdr2i8h0"></a>77.5

<a name="h.ykmyu8l6hcn2"></a>i5-19-3 [pScr]

<a name="h.3mr1227qxaog"></a>39.6

<a name="h.psdo7yxrgz7u"></a>i5-19-4 [pScr]

<a name="h.d2gmy5rmmpa1"></a>89.5

<a name="h.whyjlta6ggiw"></a>i5-19-5 [Cra]

<a name="h.nvrasbi1dfp7"></a>71.6

<a name="h.2adme0o5ap57"></a>i5-19-6 [Cra]

<a name="h.jfs9zfjou67u"></a>120.2

<a name="h.g3ggqz1i48at"></a>i5-19-7 [Com C]

<a name="h.zieop85xawgl"></a>124.8

<a name="h.al7jvioy9ey8"></a>i5-19-8 [Com C]

<a name="h.qquo1hy4hm57"></a>101.6

<a name="h.sqlxlp2zkwfk"></a>i5-19-9

<a name="h.v2yehdjyl87j"></a>108.9

<a name="h.o2jm3ibgeqzn"></a>i5-19-10

<a name="h.r9qkczavx1zz"></a>102.3

<a name="h.3sd8ogw68ews"></a>i5-19-11 [Scr]

<a name="h.t04wha5b9m3c"></a>85.8

<a name="h.toa1hyt6erb"></a>i5-19-12

<a name="h.l2uogxg89tml"></a>86.9

<a name="h.xyabvc51wqmz"></a>i5-19-13 [pFru]

<a name="h.t6elcnx8txdu"></a>101.4

<a name="h.uwy9t1l2s2f1"></a>i5-19-14

<a name="h.zgdrxg3hnv3k"></a>97.8

<a name="h.cb1zi8x4xf3h"></a>

<a name="h.podivfsbn9ht"></a>

<a href="#" name="e792f2144a5f6b55832d2e140860bcc0027fa187"></a><a href="#" name="17"></a><tbody></tbody>

<a name="h.oiv53yapn8kh"></a>Digestion

<a name="h.ou4hb7h0seu9"></a>i5-19-1

<a name="h.qte7npmt8r5c"></a>i5-19-2

<a name="h.t9s41xvjzho"></a>i5-19-3

<a name="h.40i1l3i78is1"></a>i5-19-4

<a name="h.br6zg9exyqka"></a>i5-19-5

<a name="h.ii3zqc2ors8j"></a>i5-19-6

<a name="h.fnvbqzaz7j1v"></a>i5-19-7

<a name="h.foy8blyca3l"></a>i5-19-8

<a name="h.rf3ds9ps6nzd"></a>DNA

<a name="h.zfo8ozunugrr"></a>4

<a name="h.68oznqb11yif"></a>4

<a name="h.1cme7yxrlqj6"></a>4

<a name="h.5l2uwmewg7ig"></a>4

<a name="h.58m87rj49d9u"></a>4

<a name="h.kvb2741eni0g"></a>4

<a name="h.lsdtxms32y8a"></a>4

<a name="h.c1b212wrflbg"></a>4

<a name="h.xikbro4ycyy1"></a>Buffer

<a name="h.vqsvtvp4raop"></a>1

<a name="h.jqxro38sinut"></a>1

<a name="h.2wn1xtn04gdc"></a>1

<a name="h.esnrwhx0uqms"></a>1

<a name="h.ug391abaxoi8"></a>1

<a name="h.4n9bhe83qr01"></a>1

<a name="h.v2f5x9dp0y5s"></a>1

<a name="h.905i5wpg00mj"></a>1

<a name="h.ihwa2erhrsb"></a>EcoR

<a name="h.41npynb4vjfh"></a>1

<a name="h.guc33o4xo1ny"></a>1

<a name="h.70bbc5qgkzwy"></a>1

<a name="h.m2ea79t163dd"></a>1

<a name="h.o4d1umcs55jw"></a>1

<a name="h.rl19xxze3i3y"></a>1

<a name="h.nh58fgbq8r36"></a>1

<a name="h.mi69vlcxfy8l"></a>1

<a name="h.mwpm9bcdyuz"></a>PstI

<a name="h.pbvtqamppw6t"></a>1

<a name="h.gbl8y6odfyfe"></a>1

<a name="h.8adi8qp99j8w"></a>1

<a name="h.260azt8lkiok"></a>1

<a name="h.41nqramq4vdp"></a>1

<a name="h.gad2myviufeq"></a>1

<a name="h.8cusr7ukuysz"></a>1

<a name="h.kiiioy1rzgw8"></a>1

<a name="h.41vrfxo8n7q1"></a>Water

<a name="h.dk5jox3vckg2"></a>3

<a name="h.a2vkupkprp96"></a>3

<a name="h.5dsl3xrxgwhv"></a>3

<a name="h.c860uhtzb5c"></a>3

<a name="h.iyx6d43bxlro"></a>3

<a name="h.meo2phn740du"></a>3

<a name="h.x8ehq55mafpt"></a>3

<a name="h.nw7dgbn47q03"></a>3

<a name="h.k9qik7fcg9bf"></a>Total

<a name="h.cul31bek5537"></a>10

<a name="h.f1xx1r7dzjjr"></a>10

<a name="h.ba6wq53qokt0"></a>10

<a name="h.41opoprgaair"></a>10

<a name="h.1ncbe75oart8"></a>10

<a name="h.txhacg7bmkcp"></a>10

<a name="h.ruryne7tthly"></a>10

<a name="h.ph7l6nwj0gjg"></a>10

<a name="h.7h62glaa4yxs"></a>

<a name="h.yiydd1897x9l"></a>

<a href="#" name="6e3d096d4f19e3b479acad84230ddfd1ed51fb99"></a><a href="#" name="18"></a><tbody></tbody>

<a name="h.w5r95sgzz3de"></a>Digestion #2

<a name="h.shg4jq7tk7e6"></a>1

<a name="h.i4tyvwlnt2cz"></a>2

<a name="h.4e8wcbhlw2uf"></a>3

<a name="h.yy6qr0cckv8x"></a>4

<a name="h.y2dm5eoypfl7"></a>5

<a name="h.ej3acoipgytk"></a>6

<a name="h.9c0kciymnagl"></a>7

<a name="h.waz255hd9f70"></a>8

<a name="h.25rke0gez3u"></a>DNA

<a name="h.d1o0e83sow7k"></a>4

<a name="h.g0jcy4txksbh"></a>4

<a name="h.sjk3vdrjgr0i"></a>4

<a name="h.s769fou19dqv"></a>4

<a name="h.wofmxmlpgegu"></a>4

<a name="h.490r9syon6yy"></a>4

<a name="h.uvj00hgmzve4"></a>4

<a name="h.yfgfb24q47wv"></a>4

<a name="h.yxfyh25kr"></a>Buffer

<a name="h.17jjqi4cko9w"></a>2

<a name="h.tbbryu1fptjw"></a>2

<a name="h.kil451gf68zb"></a>2

<a name="h.6bthosuis2ld"></a>2

<a name="h.5ruryk190l6y"></a>2

<a name="h.otjq8bsm9qzy"></a>2

<a name="h.p0coi219v6fu"></a>2

<a name="h.sxsz8pmiwxo4"></a>2

<a name="h.yo40h58dwy19"></a>EcoR

<a name="h.k66gxom0sihr"></a>1

<a name="h.adf4asiw7tc8"></a>1

<a name="h.3nnvtvje8bt3"></a>1

<a name="h.ml3rbpcgyxov"></a>1

<a name="h.kfp0hy18jq7t"></a>1

<a name="h.3bvz2np9788h"></a>1

<a name="h.xtmaj5gtpije"></a>1

<a name="h.oxd61apfk9qv"></a>1

<a name="h.irelmh3yosec"></a>PstI

<a name="h.s3uaas3bjpb0"></a>1

<a name="h.uo5q6iu6tjjc"></a>1

<a name="h.z9n2nm7tlijj"></a>1

<a name="h.ncu2jl8fqkt"></a>1

<a name="h.w1tglhjopah9"></a>1

<a name="h.lbrsjtd160he"></a>1

<a name="h.g7kckqak8syq"></a>1

<a name="h.4gkr4c91nlt2"></a>1

<a name="h.83pjd4a6h35b"></a>Water

<a name="h.36d3xu7uq87m"></a>12

<a name="h.8j42nnal03uu"></a>12

<a name="h.qleutk3745uj"></a>12

<a name="h.ymia2hk4ktkj"></a>12

<a name="h.xtg7amrq798x"></a>12

<a name="h.oe3428yq07hx"></a>12

<a name="h.i3rsk29jxrhj"></a>12

<a name="h.jsd4zv9zl3rh"></a>12

<a name="h.e6x30jr1n0q3"></a>Total

<a name="h.1vg74tppjqxe"></a>20

<a name="h.efr8py7hav4"></a>20

<a name="h.vz36gqyrwpjw"></a>20

<a name="h.gc3aj52yo3wh"></a>20

<a name="h.bbbs8h13lh35"></a>20

<a name="h.47r73jwcbtx4"></a>20

<a name="h.9gu3hni0qj2k"></a>20

<a name="h.rsfilo4bltd4"></a>20

<a name="h.kyc9zicsa89a"></a>

<a name="h.luqq5le1czzf"></a>8/4

<a name="h.sgg25dthi5y5"></a>

<a href="#" name="b65b6d34fbf5c5fb5aaaa4002212f34d7190e2a0"></a><a href="#" name="19"></a><tbody></tbody>

<a name="h.a5jijuql1rvr"></a>Digestion

<a name="h.ezp4ebrmrt24"></a>i5-19-9

<a name="h.iabe8cgq9hmu"></a>i5-19-10

<a name="h.7yg7kzgsqtkh"></a>i5-19-11

<a name="h.3q6b1wcbissm"></a>i5-19-12

<a name="h.k8ue8imb8tnj"></a>i5-19-13

<a name="h.ma78axahim7x"></a>i5-19-14

<a name="h.61t44pohvsaq"></a>DNA

<a name="h.x56fdk4v3ovw"></a>2

<a name="h.2hvapkiw4m87"></a>2

<a name="h.6f886gfut9m0"></a>2

<a name="h.h8oscj46xqwu"></a>2

<a name="h.kbbpg5ihzoll"></a>2

<a name="h.my8jpea79q93"></a>2

<a name="h.w9esfgelg75h"></a>Buffer

<a name="h.cfww47qpd4af"></a>1

<a name="h.qdl9k4lnp4su"></a>1

<a name="h.abiben5rcnr3"></a>1

<a name="h.3qcfojjf5lcr"></a>1

<a name="h.w7s2l12kuc50"></a>1

<a name="h.dzu83mp3t41"></a>1

<a name="h.af2xg2y7avl3"></a>EcoR

<a name="h.ni5uft1ulrul"></a>1

<a name="h.1wo6olhwpjw2"></a>1

<a name="h.wchoy4dr7x4g"></a>1

<a name="h.tbg88l1b6cy2"></a>1

<a name="h.c5y5gdkqyue2"></a>1

<a name="h.1am4iyn15gpa"></a>1

<a name="h.4tfklkehn3i5"></a>PstI

<a name="h.y5p3h0ubvj0y"></a>1

<a name="h.6i9m52boa53p"></a>1

<a name="h.13m0x4keo2uy"></a>1

<a name="h.yfvo3v5lne2u"></a>1

<a name="h.l32zngxtccv"></a>1

<a name="h.xiw4zxi2jl9m"></a>1

<a name="h.zi71ebctrsd"></a>Water

<a name="h.d0clbz3fd71r"></a>5

<a name="h.ryzpdn4j22ul"></a>5

<a name="h.gmgufotxnxut"></a>5

<a name="h.kp5bilx6wvit"></a>5

<a name="h.3vycwm7wlzip"></a>5

<a name="h.6c3d5q4bnsd1"></a>5

<a name="h.pfexr4rksv9i"></a>Total

<a name="h.xs9j65pjtnxv"></a>10

<a name="h.ayq5fs1mxbjc"></a>10

<a name="h.pk8u9by8feav"></a>10

<a name="h.aou9icqk61cv"></a>10

<a name="h.ez4mpw3kmvv0"></a>10

<a name="h.vfyzzhbb5ljt"></a>10

<a name="h.2yrznbpl1lfj"></a>

<a name="h.o6k1q9vzjd7a"></a>8/5

<a name="h.2jmyh9gl83yq"></a>Digestion

<a name="h.ii8nthcymtpb"></a>

<a href="#" name="5d3e127f652e674bf47a39df42206f4c48805de0"></a><a href="#" name="20"></a><tbody></tbody>

<a name="h.a6dix5iagyun"></a>

<a name="h.n9qgtg7uhhub"></a>i5-19-1

<a name="h.x3s5avd5kmni"></a>i5-19-3

<a name="h.4gq7ivf6m0v5"></a>i5-19-5

<a name="h.73crz0713h49"></a>i5-19-7

<a name="h.6nra6sudasj9"></a>i5-19-11

<a name="h.ao2ac858ohn1"></a>i5-19-13

<a name="h.ooejk4dkc05a"></a>DNA

<a name="h.wjvlwqn0s8dc"></a>5

<a name="h.eeg76xr9b7fv"></a>12.5

<a name="h.pfzeo3ypcsto"></a>7.1

<a name="h.qaoefpqrj4d7"></a>4

<a name="h.qu57tb5cdrok"></a>5.8

<a name="h.hgttgxoa6ab0"></a>5

<a name="h.e2razpc3mzde"></a>Buffer

<a name="h.f6uav9yzdyuy"></a>2

<a name="h.4aqyawqr5qv3"></a>2

<a name="h.b6rdcctzd01d"></a>2

<a name="h.g5zt6eo2s21p"></a>2

<a name="h.9skiq0maxx9b"></a>2

<a name="h.qk65utt0ptcm"></a>2

<a name="h.4bnct5urijfx"></a>EcoR

<a name="h.33oeogcp86tv"></a>1

<a name="h.jbie2odvtq7x"></a>1

<a name="h.xblh5q1t1us0"></a>1

<a name="h.hkms45pn6iwi"></a>1

<a name="h.3j60bs4b7afo"></a>1

<a name="h.l9onmyyqpe7z"></a>1

<a name="h.401yo1d7kmd3"></a>PstI

<a name="h.ifx7lnwalcxl"></a>1

<a name="h.bw9e2h26z0uf"></a>1

<a name="h.dnizgujui6pw"></a>1

<a name="h.x5v4qjvauhyc"></a>1

<a name="h.stev3lmw8nlg"></a>1

<a name="h.tt6o8acvxkx"></a>1

<a name="h.x6hciqcwfojq"></a>Water

<a name="h.9ht31r5zgfvd"></a>11

<a name="h.gv4kxpd8xtgw"></a>3.5

<a name="h.yp8bd29zeip0"></a>8.9

<a name="h.n372tyu3vdn2"></a>12

<a name="h.or1waswqa7s0"></a>10.2

<a name="h.26z1dl5b9lkj"></a>11

<a name="h.eho85b950qz4"></a>Total

<a name="h.yfnq9ah2mle0"></a>20

<a name="h.wqspntd4kqe0"></a>20

<a name="h.2jm6j51z1lg"></a>20

<a name="h.9q55arl12i8e"></a>20

<a name="h.wdg4qulbn5ru"></a>20

<a name="h.bgbd4xfuntjm"></a>20

<a name="h.5zjr9xt41w5m"></a>

<a href="#" name="edfa04fda6ad415d6e2fba616fd8950368ba86e6"></a><a href="#" name="21"></a><tbody></tbody>

<a name="h.a6dix5iagyun"></a>

<a name="h.n9qgtg7uhhub"></a>i5-19-1

<a name="h.ooejk4dkc05a"></a>DNA

<a name="h.wjvlwqn0s8dc"></a>5

<a name="h.e2razpc3mzde"></a>Buffer

<a name="h.f6uav9yzdyuy"></a>2

<a name="h.4bnct5urijfx"></a>EcoR

<a name="h.33oeogcp86tv"></a>1

<a name="h.401yo1d7kmd3"></a>PstI

<a name="h.ifx7lnwalcxl"></a>1

<a name="h.x6hciqcwfojq"></a>Water

<a name="h.9ht31r5zgfvd"></a>11

<a name="h.eho85b950qz4"></a>Total

<a name="h.yfnq9ah2mle0"></a>20

<a name="h.9s6iusvs6p02"></a>

<a name="h.31al3sts3g4k"></a>Picked colonies from transformation of 8/2 and 8/1

<a name="h.6koh1p1nz9b7"></a>

<a name="h.fltmgetfd1cq"></a>Colonies from 8/1

<a name="h.ap81cglnf23i"></a>1→ i5-22-1         Com B

<a name="h.17hi9mgj55xx"></a>4→ i5-22-2        pScr        Made glycerol stock for all 6

<a name="h.l5rilafwggo2"></a>5→ i5-22-3         Cra

<a name="h.4rt4e9ch9m8g"></a>7→ i5-22-4         Com C

<a name="h.ibu662kjaieq"></a>

<a name="h.mzbnml4va3h9"></a>Colonies from 8/2

<a name="h.bt2c6b5iji0t"></a>3 → i5-22-5        SCr

<a name="h.am7rhjrwfeew"></a>6→ i5-22-5        pFru

<a name="h.3osb89fv4fo1"></a>

<a name="h.n85mmvxms0xm"></a>8/6

<a name="h.uhbw6a447c9c"></a>Miniprep        Concentrations (ng/ul)

<a name="h.u1c3fa9lxf43"></a>i5-22-1                67.9        Com B

<a name="h.2s16kmp8fwjo"></a>i5-22-2                53.0        pScr

<a name="h.x2riupq5fzvg"></a>i5-22-3                90.9        Cra

<a name="h.3lehouxtgx4g"></a>i5-22-4                68.1         Com C

<a name="h.ghmduw89ynii"></a>i5-22-5                101.8        Scr

<a name="h.lj0ad7dpkd0a"></a>i5-22-6                185.5        pFru

<a name="h.tamily566yxl"></a>

<a name="h.2pzbef12bxzr"></a>Test Digestion

<a name="h.qnwry7b6xsb"></a>

<a href="#" name="810c30ac7ea7af7f46d0637a23252142e0dd376c"></a><a href="#" name="22"></a><tbody></tbody>

<a name="h.yb5evczhpuom"></a>

<a name="h.38w6pbrpqzdp"></a>i5-22-1

<a name="h.zgn0l6d6vb5t"></a>i5-22-2

<a name="h.r0yng41r0sic"></a>i5-22-3

<a name="h.p87wenjmpjwd"></a>i5-22-4

<a name="h.gkiou64ws2yz"></a>i5-22-5

<a name="h.v2g12flru3jf"></a>i5-22-6

<a name="h.va3lacp26cyj"></a>DNA

<a name="h.yg4cfww47xgo"></a>3

<a name="h.9tdm4gqjkkrq"></a>4

<a name="h.442vr9tpdgd"></a>2

<a name="h.o8mgufjbn5t0"></a>2.8

<a name="h.dt9091mp8qd3"></a>2

<a name="h.ulcrx6jqi4nf"></a>1

<a name="h.kcdjdapobt6l"></a>Cutsmart

<a name="h.elf3qphugt90"></a>2

<a name="h.3aenyfvkyno8"></a>2

<a name="h.ngwgkceddqu3"></a>2

<a name="h.vls4ubjauel4"></a>2

<a name="h.j3boni8avurg"></a>2

<a name="h.sofpjesu1oxp"></a>2

<a name="h.5ybhn2jvcnur"></a>EcoR-HF

<a name="h.85q2gdi4ggxr"></a>1

<a name="h.twfsbe5gdm9"></a>1

<a name="h.c1ss864a3wyk"></a>1

<a name="h.nmpu7yj3iswf"></a>1

<a name="h.h4x5nkmbjj16"></a>1

<a name="h.8wblq31iu8g7"></a>1

<a name="h.diecaj2350pk"></a>Spe I

<a name="h.431hxums39ls"></a>1

<a name="h.ub0o1kbmag9s"></a>1

<a name="h.bt4rjiep29yt"></a>1

<a name="h.z7prz5q61f4i"></a>1

<a name="h.5zwv242rkfvi"></a>1

<a name="h.5bb79vg9hwzd"></a>1

<a name="h.6gpw0rpjdfqg"></a>Water

<a name="h.tgamgczid50z"></a>13

<a name="h.g66iquhf4yb2"></a>12

<a name="h.daevk46emsi9"></a>14

<a name="h.5ctf9kjxkhwb"></a>13.2

<a name="h.chhppzctwrg1"></a>14

<a name="h.plsnqnkfaspd"></a>15

<a name="h.2r87y2owrqhc"></a>Total

<a name="h.cmo9e7puvjlm"></a>20

<a name="h.ag8h3mnqd2l7"></a>20

<a name="h.p4uocxac3rwe"></a>20

<a name="h.izmeh02q5kv7"></a>20

<a name="h.k5xtypjrck2a"></a>20

<a name="h.uyb7c1nwgy0u"></a>20

<a name="h.tgwmyrwiukz7"></a>

<a name="h.e4kpgf1mueqv"></a>Digestion of backbone (pLac-GFP i5-11-5)

<a name="h.stcgnprpfqqm"></a>

<a href="#" name="2ba8e1570a206c8c0cd15a29df10cc1f3dacbf85"></a><a href="#" name="23"></a><tbody></tbody>

<a name="h.d25ftzkasjyj"></a>

<a name="h.vwcs3u4w170g"></a>Com C (54.1)

<a name="h.c6xlvkt0pe77"></a>Com D (46.4)

<a name="h.o4fv70lu3990"></a>ComE (84.3)

<a name="h.44ngp9sc0gsx"></a>DNA

<a name="h.roe6ssa2ml6y"></a>40

<a name="h.fdpi22eu3m0i"></a>20

<a name="h.vg5gqoz8kjjv"></a>25

<a name="h.q0d68ztwehmr"></a>Buffer

<a name="h.v3t9mcfcbb2p"></a>5

<a name="h.dyjmwex93rs8"></a>5

<a name="h.9okzlob9xvmu"></a>5

<a name="h.nca0yzv523b"></a>Eco R

<a name="h.vh9t3cy5bmo6"></a>1

<a name="h.b6x14t79v1k4"></a>0

<a name="h.np51w6d9b29k"></a>0

<a name="h.5ago9jf3relr"></a>PstI

<a name="h.gqfk1jvl3ux7"></a>0

<a name="h.8d23ozny1z0m"></a>0

<a name="h.85nkqupm56uz"></a>1

<a name="h.tqxwofs4qxbc"></a>XbaI

<a name="h.dwkx341va5rr"></a>0

<a name="h.b43lcolhjx74"></a>1

<a name="h.5235tbrpkcom"></a>1

<a name="h.pznia856efr"></a>SpcI

<a name="h.hf4euicphgpb"></a>1

<a name="h.yu17ovg1nn2k"></a>1

<a name="h.sip7erk41zf6"></a>0

<a name="h.wnwxgm43end6"></a>Water

<a name="h.zdgligi8l7q8"></a>3

<a name="h.cukagcofwo2c"></a>23

<a name="h.xn42hpzcmfmz"></a>18

<a name="h.pvrd163duddt"></a>Total

<a name="h.i9ljeyx3xeak"></a>50

<a name="h.enwfnqcs2nbq"></a>50

<a name="h.j2h3yznwe43u"></a>50

<a name="h.s31a92d1wvih"></a>

<a name="h.y9wahg3dflx7"></a>Digestion of insert

<a name="h.ai7xrolf51jo"></a>

<a href="#" name="4cdf434ed1f855c880097220a3ba4c66edfc753a"></a><a href="#" name="24"></a><tbody></tbody>

<a name="h.uh8rvi2dflk4"></a>

<a name="h.tf68l84o8uvo"></a>Com A

<a name="h.nz13dv1nbx1n"></a>Com D

<a name="h.bnddbpky3te0"></a>Com E

<a name="h.44ngp9sc0gsx"></a>DNA

<a name="h.25pbpv9anmf5"></a>30

<a name="h.z0agsj6mnyww"></a>30

<a name="h.ssbk3twkauok"></a>30

<a name="h.q0d68ztwehmr"></a>Buffer

<a name="h.z5j53jq0ix0"></a>5

<a name="h.gbddpo6rdg2a"></a>5

<a name="h.y981msirfbq4"></a>5

<a name="h.nca0yzv523b"></a>Eco R

<a name="h.asxo1syv4hg1"></a>1

<a name="h.x71ph9ps5jlo"></a>0

<a name="h.caep0rvgrvh0"></a>0

<a name="h.5ago9jf3relr"></a>PstI

<a name="h.xxnp9vpfcyzi"></a>0

<a name="h.uh2ny991oqia"></a>0

<a name="h.ju4aq5ij8k44"></a>1

<a name="h.tqxwofs4qxbc"></a>XbaI

<a name="h.qihmsot6pl2a"></a>0

<a name="h.xtzo6ikfe7fq"></a>1

<a name="h.fje9ztr5fsb6"></a>1

<a name="h.pznia856efr"></a>SpcI

<a name="h.nxfafdazif22"></a>1

<a name="h.la1z3c7emzmo"></a>1

<a name="h.qceayaqmo7q1"></a>0

<a name="h.wnwxgm43end6"></a>Water

<a name="h.8ql6x1j0gvnk"></a>13

<a name="h.t9bpwap1kgw7"></a>13

<a name="h.7w9zite8hug9"></a>13

<a name="h.pvrd163duddt"></a>Total

<a name="h.659ckxaagez4"></a>50

<a name="h.ig2gsn60ly02"></a>50

<a name="h.ieswbupikysh"></a>50

<a name="h.wju7912zcsbi"></a>

<a name="h.db9jjmra7drh"></a>8/6

<a name="h.xq6o2p8t3ekh"></a>Constitutive promoters

<a name="h.htlcv52tnfpc"></a>

<a name="h.l6ban1o0huur"></a>

<a href="#" name="a706356d42cc6cb5710c29f11bea6cdecfddc9c2"></a><a href="#" name="25"></a><tbody></tbody>

<a name="h.wibnsic2a910"></a>CP Family member (strong)

<a name="h.6858tthab3pw"></a>BBa_J23119

<a name="h.x1z97dxzq6g7"></a>pSBIC3

<a name="h.gvhoulh69qzn"></a>2013 Kit plate 3

<a name="h.apwq70ggbn27"></a>18O

<a name="h.ints0anm9vhk"></a>i5-24-1

<a name="h.t0ps5t1x8q4l"></a>CP Family member (medium)

<a name="h.v56droy9k0d8"></a>BBa_K823014 (J23118)

<a name="h.82leoibdgaqw"></a>pSBIC3

<a name="h.ezuwrzapwbbw"></a>2013 Kit plate 1

<a name="h.3lqdqk898cbd"></a>22M

<a name="h.pek0wjix7k64"></a>i5-24-2

<a name="h.s5n956m48pp8"></a>CP Family member (weak)

<a name="h.3kc3i06t27zb"></a>BBa_K823012 (J23115)

<a name="h.jybz3h61sge6"></a>pSBIC3

<a name="h.h8fi7ukqrq2b"></a>2013 Kit plate 1

<a name="h.i92a4rin1bl2"></a>22I

<a name="h.o5q9bg9q4syf"></a>i5-24-3

<a name="h.1ncejgt5yklj"></a>

<a name="h.rycve1ih3gur"></a>Transformation

<a name="h.3jrw5j6p2sgm"></a>+ control i5-10-4 → 3 colonies

<a name="h.adynbsnxormw"></a>        i5-24-1 → 0 colonies

<a name="h.5rytshnqlc72"></a>        i5-24-2 → 25 colonies

<a name="h.k7raf6akjbw7"></a>        i5-24-3 → 15+ colonies

<a name="h.ixvvvk6gweyj"></a>

<a name="h.5tcllztdk398"></a>Gel Extraction

<a name="h.vfrtwfc8f8cd"></a>

<a href="#" name="3b9b104010f4b2e1060d38a2f95b5f5fd7fba671"></a><a href="#" name="26"></a><tbody></tbody>

<a name="h.m0esxbrsc1wj"></a>

<a name="h.i9bylpzyxh2"></a>Concentration (ng/ul)

<a name="h.ints0anm9vhk"></a>i5-24-1 Com D

<a name="h.szs7o03b7gdz"></a>17.4

<a name="h.pek0wjix7k64"></a>i5-24-2 Com E

<a name="h.ecp3xchccta6"></a>7.6

<a name="h.o5q9bg9q4syf"></a>i5-24-3 ComA

<a name="h.37benhd42xmk"></a>7.9

<a name="h.ints0anm9vhk"></a>i5-24-4 BackA

<a name="h.ia9g6gxge9v6"></a>13.3

<a name="h.pek0wjix7k64"></a>i5-24-5 BackD

<a name="h.icjpxuqk2dxn"></a>11.6

<a name="h.o5q9bg9q4syf"></a>i5-24-6 BackE

<a name="h.x6jb2cbnabnt"></a>9.6

<a name="h.a0wn8d1nez2l"></a>

<a name="h.vh0iwdhh5ns7"></a>8/7

<a name="h.o7ul5ljt2ywf"></a>Inoculate from transformation 8/6

<a name="h.2mgh77d71y1j"></a>i5-24-2 → 25+ colonies → picked 1 colony → i5-25-7

<a name="h.62ewmx28f7mt"></a>i5-24-3 → 15+ colonies → picked 1 colony → i5-25-8

<a name="h.yyhxqe7dj6zu"></a>

<a name="h.iqqfoukq2oz"></a>Com D vector backbone dephosphorilation

<a name="h.h8c9smgqz13j"></a>i5-25-5 → i5-25-9 12.5 ng/ul

<a name="h.hmejr6cbbi1s"></a>

<a name="h.8qibq2rdsosv"></a>Ligation

<a name="h.cr42wd9mm2t5"></a>

<a href="#" name="92d3062c8ee6a88c6fc20b0ac66d4fb45db13dbe"></a><a href="#" name="27"></a><tbody></tbody>

<a name="h.h7dmtux69b0h"></a>

<a name="h.75fcnjetnhp0"></a>Com A 3:1

<a name="h.ukmleeioa352"></a>Com A 0:1

<a name="h.g3ghsm5hssqk"></a>Com D 3:1

<a name="h.dnh0x7ywufq7"></a>Com D 0:1

<a name="h.tg789oqh35vb"></a>Com E 3:1

<a name="h.932dpd75v4u"></a>Com E 0:1

<a name="h.u5a3yt7rngho"></a>Insert

<a name="h.bj9gpbdmzw02"></a>10

<a name="h.u3k3id4m92o3"></a>0

<a name="h.7upxcedqi2ni"></a>6

<a name="h.ag8ouhvjt9pj"></a>0

<a name="h.xu0iinttgao0"></a>8

<a name="h.wavjmvuq0zyn"></a>0

<a name="h.a484e1vh4vez"></a>Vector

<a name="h.x5hvz11tyyru"></a>2

<a name="h.23wofbg7dk9o"></a>2

<a name="h.so5pbs3hicb"></a>5

<a name="h.3mxqleb9s8sn"></a>5

<a name="h.6zldclloa19r"></a>5

<a name="h.kgkhhly07n67"></a>5

<a name="h.fxjmuca5mbqo"></a>Buffer

<a name="h.684nr6u0aavk"></a>2

<a name="h.f29ne6k35ks0"></a>2

<a name="h.gwt9yiywu8g9"></a>2

<a name="h.9p1fp38hzmrf"></a>2

<a name="h.uw0txhnqieyp"></a>2

<a name="h.blp0k1q8ca3r"></a>2

<a name="h.eh8s8aai6ccl"></a>Ligase

<a name="h.rdx2fo9pjv50"></a>1

<a name="h.afzclqfltvst"></a>1

<a name="h.jn4ila6bgqp0"></a>1

<a name="h.niyq33tmnvsd"></a>1

<a name="h.mhdr3k2q70c4"></a>1

<a name="h.kdwsnfeco08e"></a>1

<a name="h.jhs7yvjkgipu"></a>Water

<a name="h.98uffvxft24t"></a>5

<a name="h.jx2mq28ndrhq"></a>15

<a name="h.avvmnx6rvcq7"></a>6

<a name="h.jomrcq1rkgq3"></a>12

<a name="h.2t6bgg8pq62k"></a>4

<a name="h.bk5em5zaqqk1"></a>12

<a name="h.r4gakjxirnm"></a>Total

<a name="h.b7px5p629wb2"></a>20

<a name="h.dr5wsphwugfr"></a>20

<a name="h.1jukh4az3av"></a>20

<a name="h.lv58iogydtel"></a>20

<a name="h.rs9niqua7llp"></a>20

<a name="h.kjbbjk1msd6u"></a>20

<a name="h.w7hero9cuyev"></a>

<a name="h.2j6yxbeu2tab"></a>                insert → i5-23-3                        i5-25-1                                i5-25-2

<a name="h.3o8psi3w6gw"></a>                Vector → i5-25-4                        i5-25-9                                i5-25-6

<a name="h.5yfxj9jlbxfe"></a>                1                        2                3                4                5                6

<a name="h.p6m8jq20861m"></a>

<a name="h.42bedh5ney9q"></a>Transformation

<a name="h.sn6yo85y7wf5"></a>i5-25-1 Com A 3:1 → 0 colonies

<a name="h.cl44wfisntbw"></a>i5-25-2 Com A 0:1 → 0 colonies

<a name="h.xgujaa4m9ze6"></a>i5-25-3 Com D 3:1 → 0 colonies

<a name="h.lu7vdyxwppfl"></a>i5-25-4 Com D 0:1 → 0 colonies

<a name="h.3kh89f44w1zf"></a>i5-25-9 Com E 3:1 →20 colonies → picked 3 colonies → i5-25-10

<a name="h.iehtfwff6p7f"></a>                                                → i5-25-11

<a name="h.5sydh01237my"></a>                                                → - control i5-25-12

<a name="h.vdvi3f1vqpef"></a>i5-25-6 Com E 0:1 → 0 colonies

<a name="h.7q6omna9ogct"></a>i5-11-5 + colonies → plenty colonies

<a name="h.zcyit1h53l3v"></a>

<a name="h.86c253es1cm9"></a>8/8

<a name="h.maa5grwkkatz"></a>Made glycerol stock from i5-15-7 and i5-25-8

<a name="h.ck7tfh7i7pmt"></a>Miniprep

<a href="#" name="ec89fb10a3a2f50f8dc02e3b45423d2b3675faa4"></a><a href="#" name="28"></a><tbody></tbody>

<a name="h.3jpbcjkctjf6"></a>

<a name="h.qs5drvo28rm7"></a>Concentration (ng/ul)

<a name="h.t0j5ex3dratj"></a>i5-25-7 (medium CP)

<a name="h.91gep83p0uur"></a>109.3

<a name="h.cutho5fp38n3"></a>i5-25-8 (small CP)

<a name="h.kxapg9r7ojwl"></a>116.7

<a name="h.xtxmx6x6whny"></a>

<a name="h.bvnzeytbhsfd"></a>Transformation

<a name="h.wg6vhgd3o65d"></a>i5-26-1        ComA 3:1

<a name="h.h76705w0uaeb"></a>i5-26-2        ComD 3:1

<a name="h.r92f30awsl43"></a>i5-26-3 pLac-GFP (+ control)

<a name="h.3dlfciq4c227"></a>i5-26-4 ComE 3:1

<a name="h.aj4foswu0mmv"></a>

<a name="h.s3iv9m4193k7"></a>Digestion

<a name="h.sa3hn2besxn6"></a>

<a href="#" name="0f6777305afa90a7628766949228ff86f8e7762f"></a><a href="#" name="29"></a><tbody></tbody>

<a name="h.1ybxzsmmqky1"></a>

<a name="h.cztt9ea1hy78"></a>i5-26-4

<a name="h.jxlyl7ugm2hk"></a>i5-26-5

<a name="h.12pcqlm112un"></a>i5-26-6

<a name="h.wk9jf1mpv9w8"></a>i5-26-7

<a name="h.evm95gi6jtb1"></a>DNA

<a name="h.xzp8jpql5w90"></a>30

<a name="h.43vqez2vhz75"></a>11

<a name="h.vsidxwd6b7mg"></a>20

<a name="h.yxlmuzunsphy"></a>20

<a name="h.11w80jnnzisx"></a>Buffer

<a name="h.op2rau3vwoi1"></a>5

<a name="h.u5r2g4aik22j"></a>5

<a name="h.93b535wzow5k"></a>5

<a name="h.2ohgeht0b18a"></a>5

<a name="h.48w8qfo4euj6"></a>PstI

<a name="h.5pgl0skigbuo"></a>1

<a name="h.r168jkjs4pk"></a>1

<a name="h.4djrpkwjmoyu"></a>1

<a name="h.cdxwrql0am71"></a>1

<a name="h.h6zo64dmnvbu"></a>SpcI

<a name="h.pedgoamanh83"></a>1

<a name="h.bano6kt8m2br"></a>1

<a name="h.df63q63ruwu3"></a>1

<a name="h.4rgl5cj93v1p"></a>1

<a name="h.veh91msc06en"></a>Water

<a name="h.damc93mc41p6"></a>13

<a name="h.4ss3yymvo82g"></a>32

<a name="h.7uzwotz35pb0"></a>23

<a name="h.2njg98ojeu4w"></a>23

<a name="h.iwuuy5rg7fcj"></a>Total

<a name="h.82wh9ptmr0k7"></a>50

<a name="h.lzlefu8t5gkh"></a>50

<a name="h.qy4uj2gg7gii"></a>50

<a name="h.6fj46w7i9aw9"></a>50

<a name="h.q7os7wcvo5yl"></a>

<a name="h.8oucsdmb4l50"></a>

<a href="#" name="ef197295390ef46b61c6f71de9807a81f1f2109a"></a><a href="#" name="30"></a><tbody></tbody>

<a name="h.7ohb181cvlc5"></a>

<a name="h.1ndnlsrexn6p"></a>i5-26-8

<a name="h.yvwbj81t7251"></a>i5-26-9

<a name="h.4z2y4vcanl7x"></a>i5-26-10

<a name="h.bb6yew8hwcb5"></a>DNA

<a name="h.tcf5ai7nvoqi"></a>20

<a name="h.30ln3nwv5bi0"></a>20

<a name="h.dbwm952yixl0"></a>20

<a name="h.u3c5408277v5"></a>Buffer

<a name="h.4k8p8j95ks0u"></a>5

<a name="h.asyhjt5797kh"></a>5

<a name="h.u0pcfq9fffqb"></a>5

<a name="h.tqq52md11g1r"></a>PstI

<a name="h.gqx5h1gwtiph"></a>1

<a name="h.8gr973v6w2ij"></a>1

<a name="h.j8dbj5ybpdft"></a>1

<a name="h.o2auivle7b2"></a>XbaI

<a name="h.s65sqjtdl1fv"></a>1

<a name="h.de99c0rajslu"></a>1

<a name="h.gso7obnhep95"></a>1

<a name="h.aop280y95qcn"></a>Water

<a name="h.bsebvjwtdzro"></a>23

<a name="h.o5xzj7daaev0"></a>23

<a name="h.2raz43srotx"></a>23

<a name="h.xy9f3ac7jb3n"></a>Total

<a name="h.z39vfuuiv370"></a>50

<a name="h.ce8zmsqe95xj"></a>50

<a name="h.6p64omi9z28m"></a>50

<a name="h.kamtocqtkydk"></a>

<a name="h.cpb6f9oy218h"></a>Transformation

<a name="h.82l5yurfhh46"></a>3:1 ComA

<a name="h.zc9adneqyswj"></a>3:1 ComD

<a name="h.u4z7e8tb0rrs"></a>3:1 ComE

<a name="h.ib77a1l87t48"></a>+ control (pLac-GFP)

<a name="h.eh7ds6f1gnpm"></a>

<a name="h.f00tibgergp"></a>8/9

<a name="h.kodcs5othrdp"></a>Ligation

<a name="h.z0kwt5nxopux"></a>insert: i5-26-10 Vector i5-26-5

<a name="h.9h5o97z62ydu"></a>

<a href="#" name="19791e3fc694b2233e965d69ebe788cf41afb465"></a><a href="#" name="31"></a><tbody></tbody>

<a name="h.7vluc54wk01l"></a>

<a name="h.k6mui7b6rz68"></a>3:1

<a name="h.dgvrvsc7bu0g"></a>0:1

<a name="h.oub71i2pw3n2"></a>Insert

<a name="h.amz5pvvj9q3t"></a>6.3

<a name="h.g3hp3ac60ptf"></a>0

<a name="h.ba6duxxfy39o"></a>Vector

<a name="h.60oo94a93h0b"></a>2

<a name="h.n9l5xk276hqv"></a>2

<a name="h.yly4vsjehj4"></a>Buffer

<a name="h.m9ba0nwqm5pd"></a>2

<a name="h.z71k2r76mt7f"></a>2

<a name="h.abw7saw8owjz"></a>Ligase

<a name="h.q3wgcgmtgmn4"></a>1

<a name="h.6dxg7vck2gbh"></a>1

<a name="h.erqt6icosda0"></a>Water

<a name="h.jl1ceixsbhx7"></a>8.7

<a name="h.gfwp6asmt3ku"></a>15

<a name="h.3ufubvxg739z"></a>Total

<a name="h.l6j7c6inlwdf"></a>20

<a name="h.zdtgvwkzd1kj"></a>20

<a name="h.68jcggnxe2kk"></a>                        1        2

<a name="h.xbyphphtdb9x"></a>

<a name="h.13k2y8znsdx"></a>Insert i5-26-10 Vector i5-25-4

<a name="h.1pw16fkk8vpm"></a>

<a href="#" name="d17a5321e97ae3ad32f7b3ff7218a6be7531f799"></a><a href="#" name="32"></a><tbody></tbody>

<a name="h.peun7r6p3on6"></a>

<a name="h.asit7cs2rk80"></a>3:1

<a name="h.xgvt5agishz"></a>0:1

<a name="h.oub71i2pw3n2"></a>Insert

<a name="h.xt55nt6qk32q"></a>6.6

<a name="h.3vknl45azwvf"></a>0

<a name="h.ba6duxxfy39o"></a>Vector

<a name="h.d392mwaeapid"></a>2.5

<a name="h.vt7xt8wbfrnz"></a>2.5

<a name="h.yly4vsjehj4"></a>Buffer

<a name="h.8gz1siehn7ls"></a>2

<a name="h.m5w9b5bldgpf"></a>2

<a name="h.abw7saw8owjz"></a>Ligase

<a name="h.7dv0ve66io28"></a>1

<a name="h.e8rwfes5l0e6"></a>1

<a name="h.erqt6icosda0"></a>Water

<a name="h.ygv2qmektbul"></a>7.3

<a name="h.nwq9epaajli6"></a>14.5

<a name="h.3ufubvxg739z"></a>Total

<a name="h.k4b8pbveipsm"></a>20

<a name="h.9a8kvehi89fe"></a>20

<a name="h.ssbodvtory9t"></a>                3                4

<a name="h.ml1j8fbaxfv8"></a>

<a name="h.l4rs6e81pgwk"></a>Insert i5-26-8  Vector i5-26-6

<a name="h.4xblm2trv2u8"></a>

<a href="#" name="aaf417d0d211b075e9abdd58751a67903bdd32fa"></a><a href="#" name="33"></a><tbody></tbody>

<a name="h.o14kiavtq1a6"></a>

<a name="h.2p4wl2lj83k8"></a>3:1

<a name="h.1s8hl78y6siy"></a>0:1

<a name="h.oub71i2pw3n2"></a>Insert

<a name="h.rsxvxg7ie1av"></a>4.36

<a name="h.q6l6h93wnemx"></a>0

<a name="h.ba6duxxfy39o"></a>Vector

<a name="h.xhwradv7ofd0"></a>2

<a name="h.hqbqsoydztdo"></a>2

<a name="h.yly4vsjehj4"></a>Buffer

<a name="h.5uydunqvx82s"></a>2

<a name="h.lg03ux2ir8a3"></a>2

<a name="h.abw7saw8owjz"></a>Ligase

<a name="h.963w6glbch7w"></a>1

<a name="h.avgzsiyct428"></a>1

<a name="h.erqt6icosda0"></a>Water

<a name="h.mn8l1ghufu4q"></a>10.64

<a name="h.fip39ewf67qm"></a>15

<a name="h.3ufubvxg739z"></a>Total

<a name="h.oz0oalujao1s"></a>20

<a name="h.qt6t1rqi4nbf"></a>20

<a name="h.cewwqq7q8bj1"></a>                5                6

<a name="h.4dvgy01tjgh8"></a>insert i5-26-9 Vector i5-26-6

<a name="h.dlzdlm52ue9"></a>

<a href="#" name="d04ab804a74f9748d8914ea8ae7ab6df1f55f82f"></a><a href="#" name="34"></a><tbody></tbody>

<a name="h.uk8xljkz43q0"></a>

<a name="h.rm2a91ypwctm"></a>3:1

<a name="h.rlch06w846tp"></a>0:1

<a name="h.u924bruxam85"></a>insert

<a name="h.2bhmpi28qiop"></a>2.3

<a name="h.v61xzojcboi6"></a>0

<a name="h.fdocnt4y7ji3"></a>vector

<a name="h.sjk306r40m4u"></a>2

<a name="h.se475ae81qyn"></a>2

<a name="h.t4a7k41jiwj"></a>buffer

<a name="h.o3qtri6whr8e"></a>2

<a name="h.e5p264vlll3s"></a>2

<a name="h.7cppxveawh2b"></a>ligase

<a name="h.a29fc4w34toq"></a>1

<a name="h.e99nn4f6ts9w"></a>1

<a name="h.ob97c8cj4okf"></a>water

<a name="h.ze8koa8orksh"></a>12.7

<a name="h.f77rwsefozr5"></a>15

<a name="h.1qxb72m36rfj"></a>total

<a name="h.8b6ffl6k30mh"></a>20

<a name="h.chvq5gaw562h"></a>20

<a name="h.5h8su0c8cvoe"></a>                7                8

<a name="h.3g8l51vx5mmt"></a>

<a name="h.31elyj5r49k7"></a>Transformation

<a name="h.v1z8vn6xu3il"></a>Control (w/ GFP) [i5-10-1] → i5-28-1

<a name="h.qakg30ck28x3"></a>Control (w/out GFP) [i5-22-2] → i5-28-2

<a name="h.9d158dy3dj5z"></a>

<a name="h.tz9j1wbg7mr3"></a>1 → i5-28-3

<a name="h.8il6mtefdzrt"></a>2 → i5-28-4

<a name="h.747dg3btie74"></a>3 → i5-28-5

<a name="h.9540h4uswcxa"></a>4 → i5-28-6        no growth

<a name="h.lrkw3udcpxuo"></a>5 → i5-28-7

<a name="h.58jyy6t51cy7"></a>6 → i5-28-8

<a name="h.vwu0q7nzmq1k"></a>7→ i5-28-9

<a name="h.2zsv5y78rfix"></a>8 → i5-28-10

<a name="h.hbt714lmlqv9"></a>

<a name="h.73fhqn2jb5kj"></a>Ligation

<a name="h.g3cakqv1n2j"></a>redo from 8/7

<a name="h.213inz40gxm2"></a>

<a name="h.5qyjxe42jdd2"></a>Transformation

<a name="h.te5asa59w3uc"></a>ComA 3:1

<a name="h.49y1cm5hrwwa"></a>Com A 0:1

<a name="h.4m8d55jvn825"></a>Com D 3:1

<a name="h.4afpflrq2tlb"></a>Com D 0:1

<a name="h.6s97dwn6g7av"></a>ComE 3:1

<a name="h.euuof7rbjt5n"></a>Com E 0:1

<a name="h.1e00fxu6rigc"></a>pLac-GFP as + control

<a name="h.r9jiwbxkqjp"></a>

<a name="h.8asdz9ocehq7"></a>8/10

<a name="h.9tu8znned25x"></a>ligation

<a name="h.7ggfrz5jwixt"></a>insert: i5-26-10 Vector i5-26-5

<a name="h.fs330v6uzncj"></a>

<a href="#" name="8f02dfea07dc25a4d7910b4bcdabaf0828fd7502"></a><a href="#" name="35"></a><tbody></tbody>

<a name="h.lr1ucm3je5ch"></a>

<a name="h.1tdfkaxf72hu"></a>3:1

<a name="h.irgy751q45bb"></a>0:1

<a name="h.29frdg7sf7qs"></a>insert

<a name="h.3jo10h4lfbwa"></a>6.3

<a name="h.66r7wdylz39h"></a>0

<a name="h.grelxwnto9yw"></a>vector

<a name="h.3uzirpqv6238"></a>2

<a name="h.f2tp69mzqtu8"></a>2

<a name="h.mr4s0tbd3zsx"></a>buffer

<a name="h.5d0nzx5y3dd7"></a>2

<a name="h.b7g51hyqpul1"></a>2

<a name="h.voe786qcl1rf"></a>ligase

<a name="h.q5ym06cr9hin"></a>1

<a name="h.o7p5fhdl1qx2"></a>1

<a name="h.kvnjo4exssvp"></a>water

<a name="h.xaalwyd7nrzb"></a>8.7

<a name="h.8xb8uwpml5mk"></a>15

<a name="h.rrlxvuele5sw"></a>total

<a name="h.mwokkpi7pthu"></a>20

<a name="h.xke9jmf50d4i"></a>20

<a name="h.mrcdf6e8qi1q"></a>                1                2

<a name="h.8pcb8d35m6as"></a>insert i5-26-10 vector i5-26-4

<a name="h.fs330v6uzncj"></a>

<a href="#" name="d9c4c682c5375fd0d45dd4d7f3730aaa47820b8a"></a><a href="#" name="36"></a><tbody></tbody>

<a name="h.lr1ucm3je5ch"></a>

<a name="h.1tdfkaxf72hu"></a>3:1

<a name="h.irgy751q45bb"></a>0:1

<a name="h.29frdg7sf7qs"></a>insert

<a name="h.3jo10h4lfbwa"></a>6.6

<a name="h.66r7wdylz39h"></a>0

<a name="h.grelxwnto9yw"></a>vector

<a name="h.3uzirpqv6238"></a>2.5

<a name="h.f2tp69mzqtu8"></a>2.5

<a name="h.mr4s0tbd3zsx"></a>buffer

<a name="h.5d0nzx5y3dd7"></a>2

<a name="h.b7g51hyqpul1"></a>2

<a name="h.voe786qcl1rf"></a>ligase

<a name="h.q5ym06cr9hin"></a>1

<a name="h.o7p5fhdl1qx2"></a>1

<a name="h.kvnjo4exssvp"></a>water

<a name="h.xaalwyd7nrzb"></a>7.3

<a name="h.8xb8uwpml5mk"></a>14.5

<a name="h.rrlxvuele5sw"></a>total

<a name="h.mwokkpi7pthu"></a>20

<a name="h.xke9jmf50d4i"></a>20

<a name="h.2zzkjr266vom"></a>                3                4

<a name="h.c7ej88x5dknt"></a>insert: i5-26-8 vector: i5-26-6

<a name="h.75jan7t5a5wa"></a>

<a href="#" name="fb06182ab4eca39bc376798c31660a3ff39e3dbc"></a><a href="#" name="37"></a><tbody></tbody>

<a name="h.rp0xx4ndzdnc"></a>

<a name="h.mcz4896ozeum"></a>3:1

<a name="h.vxgp54ojxuvj"></a>0:1

<a name="h.qausojphcizg"></a>insert

<a name="h.9nynadi0m07"></a>4.4

<a name="h.tqh3z8gi5eem"></a>0

<a name="h.g8daez7kpm6m"></a>vector

<a name="h.zbn5uiicfri2"></a>2

<a name="h.de9a00gs89ot"></a>2

<a name="h.qc6mrhqcnlyr"></a>buffer

<a name="h.pcgj2fa8t2fk"></a>2

<a name="h.lz8uvgragy7d"></a>2

<a name="h.nxwjrpkmzq6v"></a>ligase

<a name="h.atwez3jpqgqh"></a>1

<a name="h.knj4v3lna52d"></a>1

<a name="h.3mc1x2tvomwu"></a>water

<a name="h.yuz1fyimsmsf"></a>10.6

<a name="h.6bzfezftr4lh"></a>15

<a name="h.eqqsvyoz2beq"></a>total

<a name="h.slsnq3q69jdh"></a>20

<a name="h.za6kz0jioq5y"></a>20

<a name="h.5jjln8h6k8kb"></a>                        5                6

<a name="h.y7etqamfqm6a"></a>insert i5-26-9 Vector i5-26-6

<a name="h.j08bvg6vg5b6"></a>

<a href="#" name="2b6ea62b0c59c28b37143d040b37dd7b568cb551"></a><a href="#" name="38"></a><tbody></tbody>

<a name="h.o9c2o0qsvdh"></a>

<a name="h.xr45vdntahox"></a>3:1

<a name="h.xg71pdalpo99"></a>insert

<a name="h.9i2exop4xi3i"></a>5.1

<a name="h.tnj7o45aslz6"></a>vector

<a name="h.4jq6y5rz3ega"></a>2

<a name="h.8rl0imxjpnln"></a>buffer

<a name="h.h7ae1fba1ius"></a>2

<a name="h.3n097hgtubt9"></a>ligase

<a name="h.1h6286lbzxyx"></a>1

<a name="h.6j9jobvb4735"></a>water

<a name="h.jdxhm6vm65zt"></a>9.9

<a name="h.qf5w6k7j91ln"></a>total

<a name="h.7srltd35ouat"></a>20

<a name="h.2oy2xb6en0p2"></a>                7

<a name="h.sio52xcr0lwf"></a>8/10

<a name="h.424r9tg71t6"></a>Transformation

<a name="h.qv53featg3ve"></a>1 → i5-30-1 → 5+ colonies [pFru-RBS-GFP]

<a name="h.2mrodfqf4cj5"></a>2 → i5-30-2 → 0 colonies

<a name="h.h1ailh1sd55k"></a>3 → i5-30-3 → 5 colonies f[pScr-RBS-GFP]

<a name="h.h9xnfa7sv8gj"></a>4 → i5-30-4→ 0 colonies

<a name="h.f5rdsf7n4y3l"></a>5 → i5-30-5 → 60+ colonies [CP-Cra]

<a name="h.64iburyxf127"></a>6 → i5-30-6 → 60+ colonies

<a name="h.bi19hrunzili"></a>7 → i5-30-7 → 60+ colonies  [CP-ScR]