Team:UT Dallas/week9

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8/30

<a name="h.6va9tcl7j7lv"></a>Transformation

<a name="h.i48d2px73lur"></a>[RBS] 2013 kit 5 2M AMP Resistance → i5-54-1

<a name="h.hyr6w5yj32t1"></a>[RBS-YFP-TT] 2013 Kit 5 24G → i5-54-2

<a name="h.lu3y3kllta71"></a>+ control (carb) → i5-10-1

<a name="h.dw13sbbxr6be"></a>8/31

<a name="h.szn1xv6ganz"></a>0 colonies from 8/30 transformation

<a name="h.hcm79i2iu5em"></a>redo 8/30 transformation (different parts)

<a name="h.nsk5egtjm98r"></a>Transformation

<a name="h.9pbfxhlfz03m"></a>[RBS] 2012 kit 1 5J AMP → i5-55-1 (BBa_J61100)

<a name="h.sdv7ny5wcyos"></a>[RBS] 2012 Kit 1 2M AMP(pSB1A2) → i5-55-2 (BBa_B004)

<a name="h.2ec27w3hzamo"></a>[RBS] 2012 Kit 1 1H AMP → i5-55-3 (BBa_B0033)

<a name="h.tocu1xjvdsbo"></a>[RBS] 2013 kit 5 2M AMP → i5-55-4 (BBa_B0034)

<a name="h.vnf6ryt76ifp"></a>[RBS] 2013 kit 5 1H AMP → i5-55-5 (BBa_B0033)

<a name="h.5iigpmhmi8xz"></a>[RBS-YFP-TT] 2012 kit 124G AMP -->i5-55-6

<a name="h.72pscf40bq91"></a>(BBa_I13001) 2013 kit 3 18K Chlora → i5-55-7

<a name="h.ez21aopmr4ui"></a>+ control carb → i5-10-1

<a name="h.e14pzmbf7mct"></a>+ control chora → i5-11-5

<a name="h.8nxvlqs2cn65"></a>PCR

<a name="h.ccaiq1u0779a"></a>Redo PCR for comA

<a name="h.iciifl5d3dlq"></a>	<a name="h.y20oapoo5js"></a>comA
<a name="h.bewy8dup86iw"></a>DNA	<a name="h.ir2cxcfzq2dt"></a>2
<a name="h.bg9ysepp27ww"></a>F	<a name="h.810zk28087gl"></a>2.5
<a name="h.s5si3zki2xab"></a>R	<a name="h.x4s0oegmb42x"></a>2.5
<a name="h.jgd6bnjdskki"></a>Water	<a name="h.6uap6uctfrmn"></a>18
<a name="h.ghuaggkbop1"></a>Q5	<a name="h.clr9cztj0t1k"></a>25
<a name="h.9gckw01143a0"></a>Total	<a name="h.ci4nje5ulyhb"></a>50

<a name="h.31kzk6781izh"></a>PCR Purification

<a name="h.f3eu2zq36g79"></a>comA → i5-55-8→ 56.1

<a name="h.suazht8emft6"></a>Digestion

<a name="h.1tj5cnxnp3mf"></a>	<a name="h.1p0opyn0qd0c"></a>i5-55-8	<a name="h.3y6s5q4wh55h"></a>i5-11-5
<a name="h.mwkpgs250vw1"></a>DNA	<a name="h.epn81vjx3y1s"></a>30	<a name="h.tcvqros68n1r"></a>16
<a name="h.jhtzitigjuqj"></a>Buffer	<a name="h.rfbkeonw4uk5"></a>5	<a name="h.cd9gw27atdh9"></a>5
<a name="h.3u3tgyg9sili"></a>E	<a name="h.g7xxcbo7ehi"></a>1	<a name="h.jplzss8aa66b"></a>1
<a name="h.80xyx5lwri4a"></a>S	<a name="h.25hm74cm7bzd"></a>1	<a name="h.uwmg3732up8c"></a>1
<a name="h.otyxbrihkurv"></a>Water	<a name="h.juf3n9cnfrz1"></a>13	<a name="h.2pukd2n9kug1"></a>27
<a name="h.iqmd2qfwp9pk"></a>Total	<a name="h.kknzof8syojn"></a>50	<a name="h.cfvda558ygk7"></a>50
<a name="h.ke7ptjya7vwh"></a>	<a name="h.7z26y5i31xm2"></a>1	<a name="h.kl9njeryw3qb"></a>2

<a name="h.52jn5w8m2bn4"></a>PCR

<a name="h.iciifl5d3dlq"></a>	<a name="h.y20oapoo5js"></a>Dex
<a name="h.bewy8dup86iw"></a>DNA	<a name="h.ir2cxcfzq2dt"></a>2
<a name="h.bg9ysepp27ww"></a>F	<a name="h.810zk28087gl"></a>1
<a name="h.s5si3zki2xab"></a>R	<a name="h.x4s0oegmb42x"></a>1
<a name="h.jgd6bnjdskki"></a>Water	<a name="h.6uap6uctfrmn"></a>18
<a name="h.ghuaggkbop1"></a>Q5	<a name="h.clr9cztj0t1k"></a>25
<a name="h.9gckw01143a0"></a>Total	<a name="h.ci4nje5ulyhb"></a>50

<a name="h.oa404ofbxszp"></a>PCR Purification

<a name="h.enfamh7nfs61"></a>i5-56-1

<a name="h.cy4x1zpmmdci"></a>Gel Extraction

<a name="h.e5ji5b3wb7s6"></a>i5-55-8

<a name="h.zeu7y2hd4iah"></a>9/1

<a name="h.hftkb4myr1sl"></a>Picking colonies

<a name="h.ds9mr76ug59p"></a>+ control chlora → 20+ → picked 1 colony

<a name="h.4gkn8cc0ie5a"></a>+ control carb → 100+ → picked 1 colony

<a name="h.i3lvg6t58jkt"></a>i5-55-1 → 0 colonies

<a name="h.tlx8v6r7qocn"></a>i5-55-2 → 3+ colonies → picked 1 colony → i5-56-2 [RBS K1:2M]

<a name="h.884bjan0z5mf"></a>i5-55-3 → 3+ colonies → picked 1 colony → i5-56-3 [RBS K1:1H]

<a name="h.3hsr8opimvzr"></a>i5-55-4 → 0 colonies

<a name="h.gcd1xwsv7lrl"></a>i5-55-5 → 0 colonies

<a name="h.ddjz7to5cqfb"></a>i5-55-6 → 3+ colonies → picked 1 colony → i5-56-4 (AMP)

<a name="h.3nawnl5r56o8"></a>i5-55-7 → 3+ colonies → picked 1 colony → i5-56-5 (chlora) [RBS-YFP-TT]

<a name="h.5x8sdjnrcnt1"></a>9/2

<a name="h.typo3lkurdmj"></a>Glycerol stock & miniprep

<a name="h.w6hlycqm2mng"></a>i5-56-2 [RBS K1:2M] → 60.8

<a name="h.fk6uoxbxnn2p"></a>i5-56-3 [RBS K1:1H] → 59.4

<a name="h.b9vb18j83tiz"></a>i5-56-4 [RBS-YFP-TT (AMP)] → 81.9

<a name="h.2tw6kn35tsvr"></a>i5-56-5 [RBS-YFP-TT(Chlora)] → 103.9

<a name="h.lp1umd7kwt5e"></a>Digestion (Dex mutagenesis)

<a name="h.bi5f2s83s186"></a>	<a name="h.ruvc7nsgrs8m"></a>i5-53-1	<a name="h.oiie17pi8wfs"></a>i5-56-1
<a name="h.u9kxtq77o2qv"></a>DNA	<a name="h.3e20yak754hk"></a>20	<a name="h.uofs1vdahesp"></a>36
<a name="h.itbew2pwzj9e"></a>Buffer	<a name="h.6etw76jpgjr3"></a>5	<a name="h.svw3vxpf04by"></a>5
<a name="h.rx72xihdb1x6"></a>Afl II	<a name="h.gfmx328wwhn4"></a>2	<a name="h.99bin4qtg7ro"></a>2
<a name="h.lefzusarptif"></a>HindIII	<a name="h.h1wjogqgwtyc"></a>2	<a name="h.pp8bn2spmvc7"></a>2
<a name="h.f830lvcweazf"></a>Water	<a name="h.yx9h28v0you6"></a>21	<a name="h.h2mjpjltmt7t"></a>5
<a name="h.1nn8ii15w40q"></a>Total	<a name="h.dxz3sqxsso9l"></a>50	<a name="h.65pi2viuix8x"></a>50

<a name="h.d7v0uk4ra63f"></a>Digestion

<a name="h.kl11nycdrsed"></a>	<a name="h.jhsqiureo8z7"></a>i5-56-2	<a name="h.ho4lc09i8gor"></a>i5-56-5
<a name="h.gheinjw1o6li"></a>DNA	<a name="h.fkz0i3yo2mjh"></a>30	<a name="h.e2ln7aervggq"></a>20
<a name="h.l7b4msf7jtjq"></a>NEB Cutsmart	<a name="h.oig3zxblr7cj"></a>5	<a name="h.bh5xia4i149o"></a>5
<a name="h.ns3wrdf72ptp"></a>SpeI I	<a name="h.a5pqimtctvhu"></a>2	<a name="h.kktxizg10p9w"></a>2
<a name="h.kh6g8jb7v77i"></a>PstI	<a name="h.lzidfqx8vpr1"></a>2	<a name="h.mg9rn56t3i4m"></a>2
<a name="h.4kk8oiptji5"></a>Water	<a name="h.kp1bid9vn30l"></a>11	<a name="h.1kzt0mtrxs9p"></a>21
<a name="h.8u0yhnd4xeoh"></a>Total	<a name="h.6ygrsw9m88uz"></a>50	<a name="h.juoon6bsb601"></a>50
<a name="h.valgbajli50n"></a>	<a name="h.l6rg3vucxmxt"></a>i5-57-1	<a name="h.fbmopcq0cckp"></a>i5-57-2

<a name="h.56elclyvztzp"></a>Ligation

<a name="h.cmiy85ekg38o"></a>Insert: i5-26-8 (Fru)

<a name="h.dl1cs9rwyn8z"></a>Vector: i5-57-1 (BB-RBS)

<a name="h.oomc0irggwyi"></a>	<a name="h.qq122y5lt2gy"></a>3:01	<a name="h.aj3ttriezifm"></a>0:1
<a name="h.8drvdh2niwwb"></a>Insert	<a name="h.vwiv8ltxwsne"></a>4.4	<a name="h.m0doe69e7s7y"></a>0
<a name="h.pb9nna8fvyrq"></a>Vector	<a name="h.o247tcllwakc"></a>2.74	<a name="h.184iqvx6q4k3"></a>2.7
<a name="h.oancrb28kgt"></a>Buffer	<a name="h.e88bg7v1r88c"></a>2	<a name="h.b24fopydwyon"></a>2
<a name="h.ru4uxtis1you"></a>Ligase	<a name="h.kmfgq437hn9i"></a>2	<a name="h.vrvpkdg1mdtz"></a>2
<a name="h.lvjsmkmo83y9"></a>Water	<a name="h.1fgu1no1u5nx"></a>8.9	<a name="h.h7okwi4sqmf"></a>13.3
<a name="h.ielqmn74z12"></a>Total	<a name="h.v4ijjpggzldx"></a>20	<a name="h.6enj9bjpc5m4"></a>20
<a name="h.xs6iue3wzylh"></a>	<a name="h.8s05w3lupgwg"></a>1	<a name="h.u0vlj9fggxv2"></a>2

<a name="h.lb3fpg41vbjm"></a>Insert: i5-26-9 (scr)

<a name="h.txftv2bbjc7f"></a>Vector: i5-57-1 (BB-RBS)

<a name="h.oomc0irggwyi"></a>	<a name="h.qq122y5lt2gy"></a>3:01	<a name="h.aj3ttriezifm"></a>0:1
<a name="h.8drvdh2niwwb"></a>Insert	<a name="h.vwiv8ltxwsne"></a>5.2	<a name="h.m0doe69e7s7y"></a>0
<a name="h.pb9nna8fvyrq"></a>Vector	<a name="h.o247tcllwakc"></a>2.7	<a name="h.184iqvx6q4k3"></a>2.7
<a name="h.oancrb28kgt"></a>Buffer	<a name="h.e88bg7v1r88c"></a>2	<a name="h.b24fopydwyon"></a>2
<a name="h.ru4uxtis1you"></a>Ligase	<a name="h.kmfgq437hn9i"></a>2	<a name="h.vrvpkdg1mdtz"></a>2
<a name="h.lvjsmkmo83y9"></a>Water	<a name="h.1fgu1no1u5nx"></a>8.1	<a name="h.h7okwi4sqmf"></a>13.3
<a name="h.ielqmn74z12"></a>Total	<a name="h.v4ijjpggzldx"></a>20	<a name="h.6enj9bjpc5m4"></a>20
<a name="h.xs6iue3wzylh"></a>	<a name="h.8s05w3lupgwg"></a>3	<a name="h.u0vlj9fggxv2"></a>4

<a name="h.gp8xyzcg4rsc"></a>1-4 Need carb plates

<a name="h.kfgwziwcxlmt"></a>Insert: i5-38-7 (pFru-RBS-GFP)

<a name="h.nvb4qi7vwlqk"></a>Vector: i5-57-2 (BB-RBS-YFP-TT)

<a name="h.oomc0irggwyi"></a>	<a name="h.qq122y5lt2gy"></a>3:01	<a name="h.aj3ttriezifm"></a>0:1
<a name="h.8drvdh2niwwb"></a>Insert	<a name="h.vwiv8ltxwsne"></a>4.8	<a name="h.m0doe69e7s7y"></a>0
<a name="h.pb9nna8fvyrq"></a>Vector	<a name="h.o247tcllwakc"></a>2.4	<a name="h.184iqvx6q4k3"></a>2.4
<a name="h.oancrb28kgt"></a>Buffer	<a name="h.e88bg7v1r88c"></a>2	<a name="h.b24fopydwyon"></a>2
<a name="h.ru4uxtis1you"></a>Ligase	<a name="h.kmfgq437hn9i"></a>2	<a name="h.vrvpkdg1mdtz"></a>2
<a name="h.lvjsmkmo83y9"></a>Water	<a name="h.1fgu1no1u5nx"></a>8.8	<a name="h.h7okwi4sqmf"></a>13.6
<a name="h.ielqmn74z12"></a>Total	<a name="h.v4ijjpggzldx"></a>20	<a name="h.6enj9bjpc5m4"></a>20
<a name="h.xs6iue3wzylh"></a>	<a name="h.8s05w3lupgwg"></a>5	<a name="h.u0vlj9fggxv2"></a>6

<a name="h.56u92uj5n1rg"></a>Insert: i5-38-8 (pScr-RBS-GFP)

<a name="h.8mlbqpwvd54l"></a>Vector:i5-57-2 (BB-RBS-YFP-TT)

<a name="h.oomc0irggwyi"></a>	<a name="h.qq122y5lt2gy"></a>3:01	<a name="h.aj3ttriezifm"></a>0:1
<a name="h.8drvdh2niwwb"></a>Insert	<a name="h.vwiv8ltxwsne"></a>10.2	<a name="h.m0doe69e7s7y"></a>0
<a name="h.pb9nna8fvyrq"></a>Vector	<a name="h.o247tcllwakc"></a>2.4	<a name="h.184iqvx6q4k3"></a>2.4
<a name="h.oancrb28kgt"></a>Buffer	<a name="h.e88bg7v1r88c"></a>2	<a name="h.b24fopydwyon"></a>2
<a name="h.ru4uxtis1you"></a>Ligase	<a name="h.kmfgq437hn9i"></a>2	<a name="h.vrvpkdg1mdtz"></a>2
<a name="h.lvjsmkmo83y9"></a>Water	<a name="h.1fgu1no1u5nx"></a>3.4	<a name="h.h7okwi4sqmf"></a>13.6
<a name="h.ielqmn74z12"></a>Total	<a name="h.v4ijjpggzldx"></a>20	<a name="h.6enj9bjpc5m4"></a>20
<a name="h.xs6iue3wzylh"></a>	<a name="h.8s05w3lupgwg"></a>7	<a name="h.u0vlj9fggxv2"></a>8

<a name="h.x1pyxm26h3q5"></a>5-8 Need chlora plates

<a name="h.t7zpb09pqpqu"></a>Transformation

<a name="h.gj1k6ynmmxm1"></a>1→ i5-58-1 (3:1 BBcarb-RBS-Fru)

<a name="h.ez6jeh4cuskh"></a>2 → i5-58-2 (0:1)

<a name="h.bk9rvd7w7fil"></a>3→ i5-58-3 (3:1 BBCarb-RBS-scr)

<a name="h.daxigny6xcqs"></a>4→ i5-58-4 (0:1)

<a name="h.uz6a6tm3lfec"></a>5→ i5-58-5 (3:1 BBChlora-RBA-YFP-TT-pFru-RBS-GFP)

<a name="h.xe0z0mwaogto"></a>6→ i5-58-6 (0:1)

<a name="h.7vh74gbth6fd"></a>7→ i5-58-7 (3:1 BBChlora-RBS-YFP-TT-pScr-RBS-GFP)

<a name="h.fdqnfpom5jcl"></a>8→ i5-58-8 (0:1)

<a name="h.p356xn2ar5e5"></a>i5-10-1 → i5-58-9 (+ control carb)

<a name="h.bm65b5v0kax3"></a>i5-11-5 → i5-58-10 (+ control chlora)

<a name="h.ll4ea7lg2v73"></a>Digestion

<a name="h.p8lbx6npblg"></a>	<a name="h.gm5b797l6ps1"></a>i5-11-5
<a name="h.dy4fn9m2nhsf"></a>DNA	<a name="h.4fclie9jprvs"></a>30
<a name="h.8ckploz1a269"></a>Buffer	<a name="h.min4hupbhxyy"></a>5
<a name="h.oxe1sg4y575t"></a>E	<a name="h.n77f1fmoyu48"></a>1
<a name="h.662136d0l39b"></a>S	<a name="h.54s6ay4m5t5x"></a>1
<a name="h.m8iwx27oqi7k"></a>Water	<a name="h.28ypvj9udsdk"></a>13
<a name="h.7rtwq8viwf59"></a>Total	<a name="h.msnnjjt5ccbb"></a>50

<a name="h.i0zsdpj8c5tj"></a> Too many bands

<a name="h.sx5eesq43ma"></a>Gel Extraction → i5-59-1

<a name="h.ezz1xhxhv7ur"></a>Picked from glycerol → i5-11-5

<a name="h.z5m1bn5gq26e"></a>9/3

<a name="h.az1omzg0e4ts"></a>Miniprep/Nanodrop

<a name="h.h2v757pdvrfy"></a>i5-11-5-1 → 248.2

<a name="h.71k238uyqpp5"></a>i5-11-5-2 → 161.6

<a name="h.achz1dll2qr"></a>i5-11-5-3 → 176.9

<a name="h.hwsxqvgb3ra"></a>i5-11-5-4 → 122.5

<a name="h.7qse2fmm1gz9"></a>i5-11-5-5 → 121.6

<a name="h.uprkl260xtj7"></a>Picking colonies

<a name="h.20c21mc9j74c"></a>i5-58-1 → 200+ colonies → picked 1 → i5-59-2 BBCarb-RBS-fru

<a name="h.wzw95ryg1wov"></a>i5-58-2→ 20 colonies

<a name="h.mdlpxqx4j3e0"></a>i5-58-3→ 200+ colonies -->picked 1→ i5-59-3 BBCarb-RBS-Scr

<a name="h.cgtdxni2m81j"></a>i5-58-4→ 40 colonies

<a name="h.mez0sqklojbw"></a>i5-58-5→ 35+ colonies → picked 1 → i5-59-4 BBChlora-RBS-YFP-TT-pFru-RBS-GFP

<a name="h.krl9ev58acor"></a>i5-58-6 → 20 colonies

<a name="h.f1jljhblo6f0"></a>i5-58-7→ 75 colonies → i5-59-5 BBChlora-RBS-YFP-TT-pScr-RBS-GFP

<a name="h.nald7jy95rq4"></a>i5-58-8 → 40 colonies

<a name="h.yiaof269j4db"></a>i5-58-9 TMTC

<a name="h.cufuw0x7db4e"></a>i5-58-10 75 colonies

<a name="h.a1bkb06t4iqm"></a>Digestion (comA backbone)

<a name="h.it4qodl6ditu"></a>	<a name="h.6ku1avxo3k8"></a>i5-11-5
<a name="h.oy6n7iybjgq6"></a>DNA	<a name="h.gr6md0wrrmgg"></a>12
<a name="h.hpgyxw36b6p"></a>Buffer	<a name="h.k77qlrv7ipcu"></a>5
<a name="h.g1k2aunq1hr4"></a>Water	<a name="h.9qvtt0tf0reg"></a>31
<a name="h.pm0cc12ktdki"></a>E	<a name="h.92w2m6fs09s8"></a>1
<a name="h.vqp16qqd5v9g"></a>S	<a name="h.qf60yb80j5ei"></a>1
<a name="h.2bm8nlfd73vv"></a>Total	<a name="h.qujd19j5wntz"></a>50

<a name="h.d47ytd3jmj3t"></a>Too many bands

<a name="h.nb4d2fdm48i5"></a>Digestion (Dex mutation)

<a name="h.it4qodl6ditu"></a>	<a name="h.6ku1avxo3k8"></a>i5-53-1
<a name="h.oy6n7iybjgq6"></a>DNA	<a name="h.gr6md0wrrmgg"></a>20
<a name="h.hpgyxw36b6p"></a>Buffer	<a name="h.k77qlrv7ipcu"></a>5
<a name="h.tle3jtpt8fxi"></a>AfI II	<a name="h.9qvtt0tf0reg"></a>2
<a name="h.pm0cc12ktdki"></a>HindIII	<a name="h.92w2m6fs09s8"></a>2
<a name="h.vqp16qqd5v9g"></a>Water	<a name="h.qf60yb80j5ei"></a>21
<a name="h.2bm8nlfd73vv"></a>Total	<a name="h.qujd19j5wntz"></a>50

<a name="h.uqvg4xkv30zt"></a>Glycerol stock picks

<a name="h.e1e5jfvq4pmn"></a>i5-56-2 RBS

<a name="h.dwndeuayz0ki"></a>i5-22-1 comB

<a name="h.prhzwgu7nn5e"></a>i5-22-4 comC

<a name="h.hurnerqys6w4"></a>i5-53-1 Dex

<a name="h.dud3km1gafts"></a>Test digestion

<a name="h.f6dv5y5d2dnh"></a>	<a name="h.3sw73t2agn8y"></a>i5-30-1	<a name="h.5b7qq4hrahx"></a>i5-30-8
<a name="h.7j6yjbdncame"></a>DNA	<a name="h.u9po95wcuzhi"></a>6	<a name="h.m7vb7bpbv8ot"></a>4
<a name="h.gfpzyuw25lze"></a>Water	<a name="h.w058egfmq1yp"></a>7	<a name="h.7tgrh2dcew76"></a>9
<a name="h.lb7j2p85nkvm"></a>Buffer	<a name="h.cglai2wucdi9"></a>2	<a name="h.rwx3uxhwxg5k"></a>2
<a name="h.glpe4psvgi4p"></a>E	<a name="h.62yx8vazz88u"></a>2.5	<a name="h.1yg6ldyqwd0j"></a>2.5
<a name="h.naxjjzrwgy58"></a>S	<a name="h.osg4ln1sukcv"></a>2.5	<a name="h.eco96vx8j"></a>2.5
<a name="h.g01jkwxm0rug"></a>Total	<a name="h.nbixn8jjkmc"></a>20	<a name="h.if10715y12qd"></a>20

<a name="h.ianicfa2cyf6"></a>Digestion of chlora backbone

<a name="h.lhkkskdvds9d"></a>	<a name="h.a9om8ak1goo0"></a>i5-30-1	<a name="h.u71v5gfxcy69"></a>i5-30-8
<a name="h.6cz7904nul21"></a>DNA	<a name="h.mggkl2cv8m8b"></a>20	<a name="h.xo8qgeip92kn"></a>20
<a name="h.zbxhwlvdy02p"></a>Water	<a name="h.k8sirbtgb6gw"></a>23	<a name="h.v14m7f4hp4m1"></a>23
<a name="h.20xo8afn6hl3"></a>Buffer	<a name="h.27oz19rr005c"></a>5	<a name="h.ssj7ii9b6qv9"></a>5
<a name="h.7womkx7n0g69"></a>E	<a name="h.ypixdaqd8l5p"></a>1	<a name="h.s8tlgwlj8l2q"></a>1
<a name="h.66a3h4yruygo"></a>S	<a name="h.a7xcmlke38x7"></a>1	<a name="h.egzm6dhr6hkq"></a>1
<a name="h.y257ifcgs3d"></a>Total	<a name="h.v3blwlky27nh"></a>50	<a name="h.cqvkpl6tuq85"></a>50

<a name="h.jx1a191hu71h"></a>Gel Extraction

<a name="h.xh30nb65fauw"></a>i5-53-1 → i5-61-1→ 2.8 (throw away)

<a name="h.cudgpaykyzs4"></a>Glycerol Stock /Miniprep

<a name="h.yt6l4rprt290"></a>i5-59-2 → 166.1 [RBS-FruR]

<a name="h.u8xc6jrld2ma"></a>i5-59-3→ 128.2

<a name="h.1f9oftvuy13j"></a>i5-59-4→ 257.3

<a name="h.7lerkp796ord"></a>i5-59-5→ 140.0

<a name="h.y4m2u5e9zhwq"></a>Test Digestion

<a name="h.9bpifq2tktlr"></a>	<a name="h.v6xer83ahelg"></a>i5-59-2	<a name="h.2955qu7l29bu"></a>i5-59-3	<a name="h.ful8ge15t9if"></a>i5-59-4	<a name="h.rtf1rpiq9dy0"></a>i5-59-5
<a name="h.34hik8naamqc"></a>DNA	<a name="h.stbjqbd4krgk"></a>2	<a name="h.aboif6cnh1j7"></a>2.5	<a name="h.ap7po1yf5i2u"></a>1.5	<a name="h.m6zxnkme699f"></a>2.5
<a name="h.15hq0okz5uox"></a>Buffer	<a name="h.g5vqihb4t3i6"></a>2	<a name="h.q32hyaylcx8t"></a>2	<a name="h.la8ltd3arbeo"></a>2	<a name="h.5ope8r9bpwzi"></a>2
<a name="h.js23ha4nhjza"></a>SpeII	<a name="h.ktzozd7aghul"></a>1	<a name="h.e4x27ub51wig"></a>1	<a name="h.83nvr0u9phce"></a>1	<a name="h.rybwox49iudo"></a>1
<a name="h.8v4gmmvktagq"></a>PstI	<a name="h.vz0gsqbdd01y"></a>1	<a name="h.odli9exhus9s"></a>1	<a name="h.6oifgt215yxf"></a>1	<a name="h.l4x97qz2h4tx"></a>1
<a name="h.174xqckdcjvg"></a>Water	<a name="h.6nmd5muu5vae"></a>14	<a name="h.ctukqbeqq9rg"></a>13.5	<a name="h.iv82w9j8al2"></a>14.5	<a name="h.5px334ocgeo0"></a>13.5
<a name="h.mw189c64i7a2"></a>Total	<a name="h.pf49ngnp62dv"></a>20	<a name="h.fz6nabbo7owd"></a>20	<a name="h.16aotm7hlvft"></a>20	<a name="h.403f9qy1b348"></a>20

<a name="h.qbqzkovxl8vc"></a>Used wrong enzymes

<a name="h.aekqlquwpra2"></a>Gel Extraction (chlora backbone)

<a name="h.3e236ws3r0h9"></a>i5-61-2→ 10.8

<a name="h.p8fzpfw821is"></a>9/5

<a name="h.tk01173pvsk4"></a>Redo Test Digestion

<a name="h.oxn5t7hqovre"></a>	<a name="h.903z9xvznu2w"></a>i5-59-2	<a name="h.vv88a1r2kjvq"></a>i5-59-3	<a name="h.xovlq5cz4p8a"></a>i5-59-4	<a name="h.2bfxlrx2vun9"></a>i5-59-5
<a name="h.sn26d297p1x8"></a>DNA	<a name="h.cdss8ggw7mqb"></a>2	<a name="h.dx2dxtlvvshy"></a>2.5	<a name="h.z00486nn5gdy"></a>1.5	<a name="h.i6gfn7276ecm"></a>2.5
<a name="h.9vyzsh6ozrmd"></a>Buffer	<a name="h.27ot5orj1b6r"></a>2	<a name="h.tffswv5p8se0"></a>2	<a name="h.kjibxkea2jy5"></a>2	<a name="h.kzxuw1jwz5bj"></a>2
<a name="h.z3k789r4ak36"></a>SpeII	<a name="h.nyfpa7i0uckf"></a>1	<a name="h.j1r54n48xbu"></a>1	<a name="h.kf02xyg49yi4"></a>1	<a name="h.idxu7ma8bggu"></a>1
<a name="h.pgaqpc9w92sd"></a>PstI	<a name="h.t6t870ktxh3q"></a>1	<a name="h.qm3zgt8oij97"></a>1	<a name="h.qe39cjoucuoh"></a>1	<a name="h.1ik0mc9kdoab"></a>1
<a name="h.kr22vxbe68zz"></a>Water	<a name="h.7pbbxpeiaygk"></a>14	<a name="h.uaoz2i9y4j2s"></a>13.5	<a name="h.cbsgd06lytxn"></a>14.5	<a name="h.tgkbjzpv39xk"></a>13.5
<a name="h.oey8wl9i0xt9"></a>Total	<a name="h.e74yhaoeaji6"></a>20	<a name="h.ddq6fiseqp2d"></a>20	<a name="h.r3vifp15uhr8"></a>20	<a name="h.ljimuusibtaf"></a>20

<a name="h.nez2qo38dag"></a>Miniprep

<a name="h.t1w02qhz20qf"></a>i5-30-8→ 120.7

<a name="h.zd84tdj9hnio"></a>i5-30-8→ 118.5 [pScr-RBS-GFP]

<a name="h.qdj620f55sfc"></a>i5-30-8→ 107.6

<a name="h.cvvzrdl0dabw"></a>i5-22-1→ 125.7[comB]

<a name="h.a991i6p1qhcx"></a>i5-33-9→ 189.0[ComA]

<a name="h.enim4v639xum"></a>i5-53-1→ 164.7[Dex]

<a name="h.7u5oq51ddf59"></a>i5-56-2→ 114.4[RBS]

<a name="h.75vgedd62cs9"></a>i5-22-4→ 163.5[ComC]

<a name="h.x5tvbsfxdhfy"></a>i5-30-8→ 140.6[pScr-RBS-GFP]

<a name="h.vijtfx43nicl"></a>i5-53-1→ 263.9[Dex]

<a name="h.m3i7ouedymbo"></a>i5-56-2→ 163.8[RBS]

<a name="h.tuz8xxq8grif"></a>i5-56-2→ 75.1[RBS]

<a name="h.gvqy815oub7i"></a>i5-56-2→ 125.8[RBS]

<a name="h.var1fe6p35v1"></a>i5-56-2→ 158.4[RBS]

<a name="h.ehyqpbtrgae0"></a>Digestion

<a name="h.c0umw71v3wds"></a>	<a name="h.rz34jn7euaou"></a>i5-56-2	<a name="h.tdwqdh4kk852"></a>i5-22-1	<a name="h.bspyrg7sz7yu"></a>i5-56-2	<a name="h.ex7j0l9lmi5v"></a>i5-22-4
<a name="h.xrxqusfkjdv2"></a>DNA	<a name="h.dgeb1mv812t0"></a>20	<a name="h.h5mpiuioev7d"></a>24	<a name="h.ax0n20j0pxyi"></a>24	<a name="h.8x6jfvaz6rv6"></a>20
<a name="h.ujzo8lylch3w"></a>Buffer	<a name="h.cailpm8gxp3l"></a>5	<a name="h.262aa3menx2y"></a>5	<a name="h.4c7eiu6fi3im"></a>5	<a name="h.l3umr3d1z3ln"></a>5
<a name="h.bnkqrg9vy5r1"></a>E	<a name="h.twqdssodug83"></a>0	<a name="h.w0jfhiv9ctbv"></a>0	<a name="h.hgagm4n6tbeo"></a>0	<a name="h.7q3264az61ga"></a>0
<a name="h.949ggesa2rln"></a>P	<a name="h.hy7l7oasv1yg"></a>1	<a name="h.mkm8vmxfec3g"></a>1	<a name="h.7muu018nss41"></a>1	<a name="h.mpbrctko37e6"></a>1
<a name="h.ciw9s3qvlz8c"></a>S	<a name="h.4lsb3pd633l1"></a>1	<a name="h.6t0yb8pf5dvj"></a>0	<a name="h.pyly4ev3prb1"></a>1	<a name="h.at76yjq26tte"></a>0
<a name="h.a2qgxrw7ez3a"></a>X	<a name="h.k59hrbtxdkbe"></a>0	<a name="h.gdhctaxs3c5x"></a>1	<a name="h.mt14wgeewmcj"></a>0	<a name="h.gytlzmjel0fz"></a>1
<a name="h.g4oz5u4iofwc"></a>Water	<a name="h.rxtlzkppjg6c"></a>23	<a name="h.chl7mgx1ou0b"></a>19	<a name="h.9zqlnznczqw8"></a>19	<a name="h.y550s87xva95"></a>23
<a name="h.sesvkw72fngw"></a>	<a name="h.89mvauhjmtfm"></a>i5-64-1[RBS]	<a name="h.ni963podzmxo"></a>i5-64-2[ComB]	<a name="h.si8i7l6bhi2l"></a>i5-64-1[RBS]	<a name="h.jtf7ebs4ubb6"></a>i5-64-3 [ComC]

<a name="h.cwgbdoc568wh"></a>Digestion

<a name="h.6c6s3l4rl0us"></a>	<a name="h.byxrh63pqpfn"></a>i5-25-7(CP)
<a name="h.cuzlmcirn7ko"></a>DNA	<a name="h.4htwcphjy9b8"></a>27.4
<a name="h.ca1swrsbohgv"></a>Buffer	<a name="h.ubc1obalgkke"></a>5
<a name="h.mx3d4v45nyks"></a>S	<a name="h.hf6p64dmts17"></a>2.5
<a name="h.g3dkuql7djte"></a>P	<a name="h.3z4spsbrob4w"></a>2.5
<a name="h.tfihndtftpra"></a>Water	<a name="h.t45dvx5kyzx3"></a>12.6
<a name="h.mhiym0hv7xqs"></a>Total	<a name="h.ke7gi1l1ebov"></a>50

<a name="h.10x4uokjam2f"></a>	<a name="h.vj0a1p42mxpq"></a>i5-59-2(RBS-Fru)	<a name="h.sauxv9b198vr"></a>i5-59-3(BBS-Scr)
<a name="h.idmz9mr2lray"></a>DNA	<a name="h.qfo1a2mvnlf0"></a>18.1	<a name="h.hjx3lw9z9yt5"></a>23.4
<a name="h.rzv5aiflnk24"></a>Buffer	<a name="h.nvqn7mwzzb5v"></a>5	<a name="h.vciq9twlsvqy"></a>5
<a name="h.sma9rk5lrfst"></a>S	<a name="h.yilb9vh1pm6l"></a>2.5	<a name="h.hfrv1nq469zy"></a>2.5
<a name="h.fcj9ul45erso"></a>P	<a name="h.mo9ex882ry63"></a>2.5	<a name="h.cobn4z44xozd"></a>2.5
<a name="h.zazfhk4v0d8n"></a>Water	<a name="h.eqiv3k412bpp"></a>21.9	<a name="h.b0jerhwyaz87"></a>26.6
<a name="h.1dafjgdujwgm"></a>Total	<a name="h.zbbvo66mt425"></a>50	<a name="h.dlldvc53kusc"></a>50
<a name="h.g69p0ygdp6p2"></a>	<a name="h.tt4u7jc682hu"></a>i5-63-2	<a name="h.w6gfbef7z54r"></a>i5-63-3

<a name="h.nej4ywcohfds"></a>Gel Purification

<a name="h.w8fwndeyv4o3"></a>i5-64-1→ 45.3→ rbs (cARB) (BBA_B0034)

<a name="h.6wfn9pfr2wxq"></a>i5-64-2→ 24.5 → ComB

<a name="h.ge58wxd4k7jz"></a>i5-64-3→ 12.4 → ComC

<a name="h.d5hbe56n28fl"></a>i5-63-1→ 16.6→ Medium CP

<a name="h.dzu85b5uo14t"></a>i5-63-2→ 27.2 → RBS-FruR

<a name="h.e7iredqm8ndo"></a>i5-63-3→ 16.3→ RBS-sucR

<a name="h.kbcx90d5o7ss"></a>Ligation

<a name="h.44mv0cscryp9"></a>	<a name="h.exf5bw39k7vq"></a>comB+RBS	<a name="h.6mzy1zn11hrk"></a>comC+RBS
<a name="h.i5n7kd7zn7d6"></a>Insert	<a name="h.ex3ypdis9rsc"></a>2,0	<a name="h.6t5i11c474t0"></a>1.5,0
<a name="h.xus7gl7i12v"></a>Vector	<a name="h.ft4giicnv8xw"></a>1.5,1.5	<a name="h.llhulwyztjvv"></a>2,2
<a name="h.pvqvqk3oy7v6"></a>Buffer	<a name="h.raur0kyreq5s"></a>2,2	<a name="h.9cwmvyivf7u"></a>2,2
<a name="h.p3kbshdor8g"></a>Ligase	<a name="h.lm2wdrqk67rq"></a>1,1	<a name="h.q2myn4ui9yfz"></a>1,1
<a name="h.9ib5ckk0pl4g"></a>Water	<a name="h.ikqq8nsd32pv"></a>13.5,13.5	<a name="h.3wvzjwk0g9y9"></a>13.5,15
<a name="h.aggw3kb96udt"></a>Total	<a name="h.bqlvfmrvuozw"></a>1	<a name="h.cg67hgltfgym"></a>2

<a name="h.6qn0lccit0ft"></a>Ligation

<a name="h.b24xhrzbsrct"></a>Insert: i5-63-3 (RS-ScrR)

<a name="h.7bvx9afqqbz1"></a>Vector: i5-63-1(Chlora+CPM)

<a name="h.lslvbea26uk2"></a>	<a name="h.ebpal9gh8ajf"></a>3:01	<a name="h.xqdvq785l8yr"></a>0:01
<a name="h.fitjm021my8a"></a>Insert	<a name="h.v7vl81we5cj"></a>3.6	<a name="h.qlgppm801z7w"></a>0
<a name="h.7yh010n6j72t"></a>Vector	<a name="h.9551xivawwhz"></a>2.46	<a name="h.hhl4ceqk9mfc"></a>2.46
<a name="h.kavgu0quh5e"></a>Buffer	<a name="h.e4smtoxm57k2"></a>2	<a name="h.yrrta1uw12c6"></a>2
<a name="h.a01ce69ac5xs"></a>Ligase	<a name="h.z7th9dpgc7"></a>1	<a name="h.5eyq4hkyyocl"></a>1
<a name="h.3t050ddk5bcj"></a>Water	<a name="h.s720k6omet8b"></a>10.9	<a name="h.hft2d7kvc551"></a>14.5
<a name="h.bg0nb1tnx6u"></a>Total	<a name="h.qpylcpo1fzgx"></a>20	<a name="h.lw8lvaiw9rqp"></a>20
<a name="h.c0ym5gvcvaa5"></a>	<a name="h.8wkmgmy6wc4w"></a>1a	<a name="h.cl3ieqxkihoq"></a>2a

<a name="h.tns2ax8ebcmr"></a>Insert: i5-63-2

<a name="h.3mwbyeo1bes5"></a>Vector: i5-63-1

<a name="h.fxox0lm0iwt3"></a>	<a name="h.tmmbflqbfp84"></a>3:01
<a name="h.jhdhflsr2bfi"></a>Insert	<a name="h.9wssriocfyyq"></a>2.2
<a name="h.eavo2nlgw96g"></a>Vector	<a name="h.16pmeui1ba3p"></a>2.4
<a name="h.gn1lmmco17uo"></a>Buffer	<a name="h.9kuiefa260lp"></a>2
<a name="h.aivdxf1c1cyx"></a>Ligase	<a name="h.4a5fd14cuuoe"></a>1
<a name="h.934td23tla2u"></a>Water	<a name="h.yckws8mcltvq"></a>12.4
<a name="h.d9iokbi0jgb4"></a>Total	<a name="h.fwuxg4eza7t8"></a>20
<a name="h.1o9pb2wf80ak"></a>	<a name="h.i28gnh57ihps"></a>3a

<a name="h.oewfve66vsx3"></a>Repressor System

<a name="h.whs62anstrrv"></a>1→ i5-65-1→ 0 colonies

<a name="h.urphfl6060am"></a>2→ i5-65-2→ 0 colonies

<a name="h.yo4k96k082iq"></a>3→ i5-65-3→ 0 colonies

<a name="h.hcbj1eayp4e"></a>comB+RBS

<a name="h.3xzj3hv8ruwf"></a>1→ i5-65-4→ 200 colonies → picked 2 → i5-65-11 & i5-65-12

<a name="h.perkmtr72bt8"></a>2→ i5-65-5

<a name="h.ki6qcida0w2c"></a>ComC+RBS

<a name="h.hob59as39tww"></a>3→ i5-65-6→ 20 colonies → picked 2→ i5-65-13 &i5-65-14

<a name="h.fvmr4v32jwua"></a>4→ i5-65-7

<a name="h.norrasbwqlt0"></a>From glycerol

<a name="h.ehv55w8w718f"></a>i5-25-7 (CPM) → i5-65-8→ infinite colonies → picked 1→ i5-65-17

<a name="h.ngss1opx69z6"></a>i5-30-1 (pFru-RBS-GFP)--> i5-65-9→ infinite colonies → i565-18

<a name="h.ch3b6d2h2l3g"></a>TT(BBa_B0015) 2013 Kit3-4F (Chlora)--> i5-65-10→ 6 colonies→ 2 picked → i5-65-19 & i5-65-20