Team:UT Dallas/week5

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7/28/2013

<a name="h.1w7xzdr4u63c"></a>PCR

<a name="h.l98dou3iv8o"></a>Primers	<a name="h.iylmgzh0plu0"></a>1	<a name="h.h7cygypzygvd"></a>2	<a name="h.htgvzlr3zeg1"></a>3	<a name="h.c9zuyf785t1k"></a>4	<a name="h.atfwyo73js4d"></a>5	<a name="h.h7dqlwqgya0e"></a>6	<a name="h.6ekecfdp08zm"></a>7	<a name="h.emgt12bv0g4g"></a>8	<a name="h.cmpkt9yv0iat"></a>9	<a name="h.je7t7q6tv1rf"></a>10
<a name="h.s8aq2lbk47y3"></a>DNA	<a name="h.rwp11tgj4ahh"></a>1	<a name="h.z2tat31a3gar"></a>1	<a name="h.i98vrbkqsqnm"></a>1	<a name="h.usa99qhtlxql"></a>1	<a name="h.yk37pz4wnolg"></a>1	<a name="h.93mvjlyrb5jn"></a>1	<a name="h.dz2ctq9w2aqk"></a>1	<a name="h.sotzy92rv1s"></a>1	<a name="h.li3a7e2bdvn1"></a>1	<a name="h.t6ryvfn516e3"></a>1
<a name="h.e6r7lvtrxib4"></a>F	<a name="h.9c8jtdfzcapw"></a>1	<a name="h.glq9z4l3ebu"></a>1	<a name="h.cy5bp9k13ic0"></a>1	<a name="h.9d7e08bmzv1d"></a>1	<a name="h.gduu3qbma75b"></a>1	<a name="h.z6t16hk7if5r"></a>1	<a name="h.39dyx564wgqh"></a>1	<a name="h.40oje46pms7d"></a>1	<a name="h.np1vcv5u8g5k"></a>1	<a name="h.emqtdc1e10t6"></a>1
<a name="h.azwizg70184b"></a>R	<a name="h.o5hmje4g5c8u"></a>1	<a name="h.ukf167shdgjo"></a>1	<a name="h.o41h7oavaiam"></a>1	<a name="h.tf8kg14l1y3r"></a>1	<a name="h.irnniwmsayy0"></a>1	<a name="h.ygpgl6h0jhre"></a>1	<a name="h.i71phhtxr5lq"></a>1	<a name="h.o1o13ry40x9f"></a>1	<a name="h.752xc4awlv9t"></a>1	<a name="h.s82d9wj9hh0f"></a>1
<a name="h.wwgb8rb2caqx"></a>Phusion	<a name="h.ot6w07vgsmho"></a>10	<a name="h.j0qp0j8yylz"></a>10	<a name="h.9ka3k0y4zl04"></a>10	<a name="h.tf6hvlrn1u79"></a>10	<a name="h.pnybwmk1amlp"></a>10	<a name="h.vyeifkgxpagy"></a>10	<a name="h.vy8zdbxwo69v"></a>10	<a name="h.13lc0p7rp4g"></a>10	<a name="h.9rvgw4inrhql"></a>10	<a name="h.bgvpr1ym61fz"></a>10
<a name="h.zfb5f4i9rdii"></a>Water	<a name="h.sta2t5jqzrvw"></a>7	<a name="h.icutzgtuflsq"></a>7	<a name="h.dvu38r3f8g1t"></a>7	<a name="h.62umehxctlvk"></a>7	<a name="h.1zld32o1mbx4"></a>7	<a name="h.q7dclgac2akm"></a>7	<a name="h.e53dqcg8lwdh"></a>7	<a name="h.thms7q28qo7y"></a>7	<a name="h.5hwyopy6g198"></a>7	<a name="h.h268jsv0xfi7"></a>7

<a name="h.625habuoaedf"></a>Digestion

<a name="h.537gplby40eo"></a>	<a name="h.xqc0lppe6krv"></a>5 (i5-13-9)	<a name="h.ubh03cwp1deq"></a>6 (i3-13-10)	<a name="h.o2wbmaardiau"></a>7 (i3-13-11)	<a name="h.ovwu3kvq8nyk"></a>8 (i3-13-12)	<a name="h.y6s4vn5x3iav"></a>9 (i3-13-13)	<a name="h.smdjo587yn7d"></a>i5-10-3 (i5-13-14)
<a name="h.xntrywweu8of"></a>DNA	<a name="h.lv3064b3lmj"></a>30	<a name="h.8ph34t10kt1k"></a>30	<a name="h.rsoug65yaof9"></a>30	<a name="h.er67n63cg2v7"></a>30	<a name="h.bqbab170861d"></a>30	<a name="h.igemoh6tuopz"></a>30
<a name="h.3z2okkavkzgn"></a>NE Buffer	<a name="h.23pmrd3j3kfz"></a>5	<a name="h.vsrsr7pdqtt0"></a>5	<a name="h.epa3h1153om3"></a>5	<a name="h.4ikn46vluxyw"></a>5	<a name="h.gxe844eltiua"></a>5	<a name="h.iaz8nua1eyfu"></a>5
<a name="h.2uuz4e40isx1"></a>EcoRI	<a name="h.i0xp596p5fw1"></a>1	<a name="h.sqlcqmgvj173"></a>1	<a name="h.cq48ztpsv71d"></a>1	<a name="h.ck15m3arqdrd"></a>1	<a name="h.93tkn3hqbi9w"></a>1	<a name="h.w0glvf9ljf32"></a>1
<a name="h.77qgyz3qlwhs"></a>Prst I	<a name="h.dotv3dly1w9n"></a>1	<a name="h.m0cauz10idxo"></a>1	<a name="h.pzaip3q8vyky"></a>1	<a name="h.5ccfp79x0sya"></a>1	<a name="h.p1vfow1jids1"></a>1	<a name="h.mq5gkij39x1m"></a>1
<a name="h.tofmw9c4te4v"></a>Water	<a name="h.6dwq0bt714dx"></a>13	<a name="h.t6qsxr11k62i"></a>13	<a name="h.dbq1oskgtj8a"></a>13	<a name="h.j6u49lawj21h"></a>13	<a name="h.loge284qwxle"></a>13	<a name="h.vmjjkyo4l4vt"></a>13

<a name="h.kvb5pahdagn6"></a>7/24/2013

<a name="h.d8tsh5kkz4h0"></a>PCR

<a name="h.q47n76xr18ri"></a>Primers	<a name="h.7rkpqmb3uwac"></a>1	<a name="h.8kvhh4ls95g1"></a>2	<a name="h.l6ktdui81p2i"></a>3	<a name="h.wg2w0swmphbf"></a>4
<a name="h.aupmsss8kqaz"></a>DNA	<a name="h.us5i9ie2ief7"></a>1	<a name="h.d1arncc0oq0i"></a>1	<a name="h.wx2lt1gmgejp"></a>1	<a name="h.moqlznfx8kxv"></a>1
<a name="h.ea6xpevky1n5"></a>R	<a name="h.2lo40l8nn351"></a>1	<a name="h.c4nvgln2gort"></a>1	<a name="h.l4ns7ijsrf9a"></a>1	<a name="h.v67x0t1gynjr"></a>1
<a name="h.g728ofbrsyd2"></a>F	<a name="h.log4kdfc2xut"></a>1	<a name="h.2tilhu3yk376"></a>1	<a name="h.mrnv1jd0m6vk"></a>1	<a name="h.19fqyxi4lj5b"></a>1
<a name="h.wgb3khr5skw2"></a>Phusion	<a name="h.63er7ixlb9ud"></a>10	<a name="h.3mxustbm97sx"></a>10	<a name="h.katv3qd44lkw"></a>10	<a name="h.q7gjuhvs0um"></a>10
<a name="h.4hdp70vtlra"></a>Water	<a name="h.sndqj4z3vcvl"></a>7	<a name="h.1f065lbzz1z"></a>7	<a name="h.gvon0ul75w87"></a>7	<a name="h.xoffqy49dsnd"></a>7

<a name="h.koawdqsmnyoh"></a>1 i5-14-1 33.3 ng/ul (Com C)

<a name="h.fv1vt78mg7ru"></a>2 i5-14-2 47.5 ng/ul (Com D)

<a name="h.3x1x0benh4pe"></a>3 i5-14-3 38.4 ng/ul (Com E)

<a name="h.4z3kd4o6avp1"></a>4 i5-14-4 27.8 ng/ul (Com A)

<a name="h.byev1ekaa63a"></a>7/30/2013

<a name="h.hhq1lr5p0okp"></a>Digestion

<a name="h.rz8lt1ph5jxn"></a>1 (i5-14-1) (Com C)

<a name="h.n0uh9rz1mf4q"></a>DNA 30 (0 left)

<a name="h.kneof781dyz1"></a>NEBuffer 5

<a name="h.i4qtg9crqsm6"></a>EcoRI 1

<a name="h.db59aiebk0lj"></a>PstI 1

<a name="h.fw9odm6ololp"></a>Water 13

<a name="h.ut30mxfqsmhf"></a>→ new i5-14-1

<a name="h.qyltrisqlr82"></a>Gel extraction i5-14-1

<a name="h.mvk988hr22kv"></a>i5-13-9 (Com B) 8.6 ng/ul

<a name="h.dj3ugc0biey"></a>i5-13-10 (ScrR) 12 ng/ul

<a name="h.5qit7p46px2g"></a>i513-11(pScr) 14.4 ng/ul

<a name="h.3do6oe7lvk2u"></a>i5-13-12 (Cra) 15.1 ng/ul

<a name="h.5y516xtktq34"></a>i5-13-13(pFru) 11 ng/ul

<a name="h.4boqh7b34zdl"></a>i5-13-14(Chlora backbone) 24 ng/ul

<a name="h.s5lp4mbltzhy"></a>i5-15-1 (Com C) 13.9 ng/ul

<a name="h.ut6ozfvulmxz"></a>Ligation

<a name="h.pfehlplbqkke"></a>Vector i5-13-14 (Chlora)

<a name="h.rwclbd7ju5o6"></a>Insert:

<a name="h.eapxxn7uc1g6"></a>i5-13-9

<a name="h.8yh49v9ynpyf"></a>i5-13-9

<a name="h.z9qy3zix2fy9"></a>i5-13-10

<a name="h.pjoqdlvv3lkm"></a>i5-13-10

<a name="h.wq1gnmk3rbzf"></a>i5-13-11

<a name="h.n8s28blj6ch8"></a>i5-13-11

<a name="h.vzyxdfb5t3sd"></a>i5-13-12

<a name="h.2qrdexvawwpk"></a>i5-13-12

<a name="h.z6v5lw64ahad"></a>i5-13-13

<a name="h.y5i9fqvu88i6"></a>i5-13-13

<a name="h.ucy77na90940"></a>i5-15-1

<a name="h.n4drv21h8wzz"></a>i5-15-1

<a name="h.4v3arya9n04w"></a>3:1

<a name="h.lze3kxcsc4ks"></a>0:1

<a name="h.7gpyhvx1ddwn"></a>3:1

<a name="h.tb20iaswej"></a>0:1

<a name="h.hf9fz1ks5wzs"></a>3:1

<a name="h.yvfgmo49urdc"></a>0:1

<a name="h.xbpyfe5ghwhd"></a>3:1

<a name="h.s365vqt413uu"></a>0:1

<a name="h.tj1zd9fyq725"></a>3:1

<a name="h.gv8xkigvkbxs"></a>0:1

<a name="h.bn78bbm9lzz5"></a>3:1

<a name="h.11iltw2g7gja"></a>0:1

<a name="h.8xwue3hhxoj2"></a>Vector

<a name="h.ueapcumgr70z"></a>2

<a name="h.a90678jyza68"></a>2

<a name="h.e6e899jjwqh"></a>2

<a name="h.v9uy9dru9lha"></a>2

<a name="h.7s651ehvei4b"></a>2

<a name="h.i107go106z3z"></a>2

<a name="h.xhtik8jfr5t8"></a>2

<a name="h.7jycjrl59rkh"></a>2

<a name="h.bw4ew4c4843j"></a>2

<a name="h.1yf4govsdmmz"></a>2

<a name="h.dy0o6cu8osv8"></a>2

<a name="h.e1bg38dvm6ts"></a>2

<a name="h.cykgsr2weglr"></a>insert

<a name="h.eizukidl4cyc"></a>7.5

<a name="h.kog90q6bx275"></a>0

<a name="h.98vnva3vgr2u"></a>6

<a name="h.9u7gxwmfqfpt"></a>0

<a name="h.azeiwoyo9gg3"></a>1

<a name="h.shdnosdzcw16"></a>0

<a name="h.xhdyltdzx6ea"></a>5

<a name="h.qb978w6jidz"></a>0

<a name="h.mdvwwm8w3gr2"></a>1.5

<a name="h.berxsw3ctoj9"></a>0

<a name="h.bws8wghda8eq"></a>1

<a name="h.exx4y5x68751"></a>0

<a name="h.ufw4a7mv08qf"></a>buffer

<a name="h.e82ys14lie5g"></a>2

<a name="h.w3awm2jc8x62"></a>2

<a name="h.ip79buro7z35"></a>2

<a name="h.2jwvv8pad0y8"></a>2

<a name="h.eui2glumbma9"></a>2

<a name="h.ij6a4wggvhr8"></a>2

<a name="h.5erup0qxiaoa"></a>2

<a name="h.34drqu9lsiip"></a>2

<a name="h.sxt14q53zna"></a>2

<a name="h.zeib64cg79d2"></a>2

<a name="h.w76ejfa0vgam"></a>2

<a name="h.sli6mh394kb5"></a>2

<a name="h.v35ueq5cr2ki"></a>ligase

<a name="h.jp2k3gq27m9p"></a>1

<a name="h.l2ezm01epi51"></a>1

<a name="h.ep4rxobcrnm0"></a>1

<a name="h.gr1lj3ltfo95"></a>1

<a name="h.gf8vy915vn"></a>1

<a name="h.trcwr9gqzc95"></a>1

<a name="h.ap4irlbntl3b"></a>1

<a name="h.brtf4pa8s3tg"></a>1

<a name="h.56a01qf31wqd"></a>1

<a name="h.8nysqqn9xpga"></a>1

<a name="h.cgwytf5eesu9"></a>1

<a name="h.b39j5p6tch1k"></a>1

<a name="h.kf7kmjciwkpj"></a>water

<a name="h.l5mxf8eibg1"></a>7.5

<a name="h.4hvglti0oxpi"></a>15

<a name="h.jr1bst19a9wz"></a>9

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

<a name="h.pndyfze6qn5i"></a>14

<a name="h.gub9i3ohcehi"></a>15

<a name="h.i2kexdv8pq33"></a>10

<a name="h.o7nzrkza94cd"></a>15

<a name="h.xpe8r41l59s0"></a>13.5

<a name="h.55gekl6q1m72"></a>15

<a name="h.ym8dfmbzpnfk"></a>14

<a name="h.1nuuijugz687"></a>15

<a name="h.xt2svhv3b8l4"></a>total

<a name="h.2tn6w1uqmqxz"></a>20

<a name="h.sdpcuzjyl7ts"></a>20

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

<a name="h.bsdbvy1o7wln"></a>20

<a name="h.jwaa3xuxas9"></a>20

<a name="h.hqmcbdgpw0lx"></a>20

<a name="h.e0ehwrwd4agz"></a>20

<a name="h.zfiuqf321edu"></a>20

<a name="h.ps0ewmqtqv9o"></a>20

<a name="h.n6pyhtm59dbi"></a>20

<a name="h.y4lt0enlcu3u"></a>20

<a name="h.riegvhuvgl4n"></a>20

<a name="h.eue2n4kohh0l"></a>1

<a name="h.ef50nw1y7qha"></a>2

<a name="h.2l6f7excv42h"></a>3

<a name="h.3c6fmyyunrm7"></a>4

<a name="h.l5ht1kgx2r3d"></a>5

<a name="h.zi9axt96eqt2"></a>6

<a name="h.qr6e4yv2ily7"></a>7

<a name="h.exxb2fxp0zbw"></a>8

<a name="h.kuz4x1yluh03"></a>9

<a name="h.7luzyy8w7tqz"></a>10

<a name="h.wzwxx8s2jhfe"></a>11

<a name="h.c4dyb9ecvegy"></a>12

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

<a name="h.i3wtjhdwzin3"></a>1

<a name="h.yqiuna5qzuv"></a>2

<a name="h.6506ohlh2qwn"></a>3

<a name="h.ic0zbvoiwg54"></a>4

<a name="h.7jw4qtpuq8u"></a>5

<a name="h.vxoc92u740ec"></a>6

<a name="h.xw9bu3ftnlrz"></a>7

<a name="h.wz32he1cowmw"></a>8

<a name="h.xu12aug1fjsk"></a>9

<a name="h.6nxy36gth9ap"></a>10

<a name="h.6vj90p5kdqyh"></a>11

<a name="h.2ivfpv951rq5"></a>12

<a name="h.xrfzapxgozv0"></a>+ control → i5-10-4

<a name="h.2sye30drygjv"></a>Inconsistent growth (repeat)

<a name="h.qdwpmlar9fxp"></a>7/30

<a name="h.h2yrqpht6uek"></a>Transformation

<a name="h.g16bsupb0if7"></a>1

<a name="h.yepbrmq46j8j"></a>2

<a name="h.yuzf97e1ycl1"></a>3

<a name="h.4k2gx4fcrj7c"></a>4

<a name="h.ytz71xaxz9i"></a>5

<a name="h.y9wrgq1sinlr"></a>6

<a name="h.f4nyxt67c1g6"></a>7

<a name="h.chgf8ey2p72w"></a>8

<a name="h.6yor6rdyrcfh"></a>9

<a name="h.ysyhpy20dxld"></a>10

<a name="h.wgqbqis3fu9k"></a>11

<a name="h.g2va1il9rme3"></a>12

<a name="h.va67wmeyjr7z"></a>+ control → i5-10-4

<a name="h.lyf956klvtnq"></a>Inoculate

<a name="h.cu4kdzhfgn9j"></a>From glycerol stock i5-11-5 (pLac-GFP)

<a name="h.cvxy2odcnx2"></a>100.4 ng/ul

<a name="h.72910tlji9sm"></a>96.4 ng/ul [pLac-GFP]

<a name="h.gxyw4fotd64k"></a>84.3 ng/ul

<a name="h.1sjg3n4ze7nq"></a>7/31

<a name="h.iwchhdbervw9"></a>Transformation Results

<a name="h.5atic82knicq"></a>1 [Com B] → 8 colonies → Picked 3 colonies→ i5-16-1

<a name="h.k4hslldhwd8r"></a> i5-16-2

<a name="h.16gg4u89efie"></a> i5-16-3

<a name="h.9m4le8otw8qu"></a>2→ 50 colonies

<a name="h.tbck2neavtxc"></a>3 [Scr] → 11 colonies → Picked 3 colonies → i5-16-4

<a name="h.orwfvyt5ue2v"></a> i5-16-5

<a name="h.242xprdzg0b5"></a> i5-16-6

<a name="h.8faqzlhxo0iq"></a>4 → 20 colonies

<a name="h.ds7rd5ef53tw"></a>5 [pScr] → 25 colonies → picked 3 colonies → i5-16-7

<a name="h.uninw76k2wf"></a> i5-16-8

<a name="h.qpdj9aujpd2t"></a> i5-16-9

<a name="h.405vih9kuvu"></a>6 → 0 colonies

<a name="h.ox6mwhqz0f0f"></a>7 [Cra] → 50+ colonies → picked 3 colonies → i5-16-10

<a name="h.3pqoz5468f0a"></a> i5-16-11

<a name="h.g56vdta42xvu"></a> i5-16-12

<a name="h.uf67ey9w9f40"></a>8 → 75+ colonies

<a name="h.aoggldrolu7c"></a>9 [pFru] → 75+ colonies → picked 3 colonies → i5-16-13

<a name="h.z6au5v2jb975"></a> i5-16-14

<a name="h.90uwnto7k70s"></a> i5-16-15

<a name="h.n89z936984bi"></a>10 → 11 colonies

<a name="h.283avhppju1z"></a>11 [Com C] → 30 colonies → picked 3 colonies→ i5-16-16

<a name="h.92qdy4j90bwo"></a> i5-16-17

<a name="h.wsv606ce7ery"></a> i5-16-18

<a name="h.ubmjrqntcnja"></a>12→ 30 colonies

<a name="h.mj4433clon1w"></a>13→ 7 colonies

<a name="h.u0ea7xjc6274"></a>14 →2 colonies

<a name="h.d9vx3nyuimz7"></a>15 → 20 colonies

<a name="h.f2cq7ihd00rt"></a>16→ 10 colonies

<a name="h.9cfq1743wgqc"></a>17 → 0 colonies

<a name="h.okzp12kpgfhd"></a>18 → 5 colonies

<a name="h.m6j8vq7xdy0l"></a>+ control → ∞ colonies

<a name="h.hkvp1ly7bxwx"></a>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.s0shikkagflk"></a>Concentrations too low

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

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

<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.ak8khbywm050"></a>Digestion

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

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

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

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

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

<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.vdlf9hfxs3za"></a>8/1

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

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

<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.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.hupfbsd3ivci"></a>8/3

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

<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.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.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