Team:UT Dallas/week4

From 2013.igem.org

Revision as of 20:53, 23 September 2013 by Alecspin (Talk | contribs)
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

7/22/2013

[GFP] – I3-5-8 Carb Resistance

[pLac]- I5-6-3 Chlora

I4-45-2 Chlora

7/23/2013

Transformation Results:

<a href="#" name="561d1c16c2e9d617f036d365af80fddf4eda0c9f"></a><a href="#" name="5"></a><tbody></tbody>

# of colonies

NEB

Commercial

I4-45-2

100+

0

I5-6-3 [pLac]

100+

90+

GFP-I3-5-8

100+

Tmtc

[pLac]-I5-6-3 → [NEB] 100+ colonies →  picked two colonies →  i5-10-3

                                                              → i5-10-4

[GFP]-I3-5-8 → [NEB] 100+ colonies → picked two colonies →  i5-10-1

                                                            →  i5-10-2

7/24/2013

Miniprep

i5-10-1[GFP]        98.4 ng/ul

i5-10-2[GFP]        61.4 ng/ul

i5-10-3[pLac]        46.9 ng/ul

i5-10-4[pLac]        61.8 ng/ul

Transformation

NEB cells

i5-6-3 (positive control) → i5-10-1 → 20 + colonies

3:1 → i5-10-2 [pLac-GFP] → 2 colonies

0:1 (negative control) → i5-10-3 [pLAc vector] → 0 colonies

7/25/2013

DNA Isolation

MGI655 → i5-11-1        34.8 ng/ul

        → i5-11-2        60.7 ng/ul

        → i5-11-3        15.5 ng/ul

        → i5-11-4        35.1 ng/ul

Enzymatic lysis buffer:

20 ul Tris-EDTA

12 ul Triton

912 ul Water

______________

56 ul lysozyme (add right before use)

7/26/2013

Glycerol Stock

i5-10-4 → i5-11-5

i5-10-5 → i5-11-6

Miniprep

i5-10-4 → i5-11-5        54.7 ng/ul [pLac-RBS-GFP]

i5-10-5 → i5-11-6        49.2 ng/ul [pLac-RBS-GFP]

PCR Reaction (MGI655)

<a href="#" name="fb2bbd567864d0e2cdd50e7c82681985e2e636d5"></a><a href="#" name="6"></a><tbody></tbody>

8

9

DNA

1

1

Primer (8F)

1

R 1

Primer (8R)

1

F 1

Polymerase (pFusion)

10

10

H₂O

7

7

Total

20

20

DNA Isolation with RNeasy Blood  and Tissue Kit

S.Mutans -->

i5-12-1 → 88.3 ng/ul

i5-12-2 → 30.3 ng/ul

7/27/2012

<a name="h.c74wfbk0ki91"></a>

<a href="#" name="568664dec79d8d575aaf1cec83403fa8c1a3ad50"></a><a href="#" name="7"></a><tbody></tbody>

<a name="h.x80mk69q957f"></a>Primer

<a name="h.lbc1tyn3yika"></a>1

<a name="h.kpjc1dskz0r9"></a>2

<a name="h.hw3zrhyk2pdl"></a>3

<a name="h.jxlb5edmq6li"></a>4

<a name="h.det2fdtn1z2t"></a>5

<a name="h.q7u5n4r5fca7"></a>6

<a name="h.vi3hb65hn03k"></a>7

<a name="h.muyiuvqm8zg"></a>8

<a name="h.mm81mm7tll0x"></a>9

<a name="h.kfwgk9phgoud"></a>10

<a name="h.kasipbw7l7q"></a>DNA

<a name="h.i9dwvpq5noho"></a>1

<a name="h.61vxj34hxkvb"></a>1

<a name="h.1zfxoe8f692d"></a>1

<a name="h.dbeix02xwbep"></a>1

<a name="h.ftxj9rii53uz"></a>1

<a name="h.713p8m2nv76l"></a>1

<a name="h.ofblf8t9axf8"></a>1

<a name="h.k698esb6eeli"></a>1

<a name="h.ml5firq4a49"></a>1

<a name="h.5osr8pdtzuqs"></a>1

<a name="h.cw0pwuwumr28"></a>Forward

<a name="h.zfc42kp5da47"></a>1

<a name="h.ac7j131q4uia"></a>1

<a name="h.mrl9catt06o5"></a>1

<a name="h.ox5es01oni9k"></a>1

<a name="h.5snlje483gyo"></a>1

<a name="h.kgcuw9be9o5v"></a>1

<a name="h.mbrwokdfb7f5"></a>1

<a name="h.ju7eg3tn6xp"></a>1

<a name="h.vk1we9vrmzyt"></a>1

<a name="h.8aerfpmc3ewm"></a>1

<a name="h.lvt2xtuz7nfx"></a>Reverse

<a name="h.rg728du8xg5"></a>1

<a name="h.71nik2lgl8ex"></a>1

<a name="h.n41bhxtl2mgo"></a>1

<a name="h.vbdtmg15wtbf"></a>1

<a name="h.qgdjpccu5y6c"></a>1

<a name="h.npc4cn3yr3es"></a>1

<a name="h.c6bwc5h8m4nk"></a>1

<a name="h.sn4clqi8f1n0"></a>1

<a name="h.y6ptei6bjx3f"></a>1

<a name="h.m2nwg1rustsy"></a>1

<a name="h.sq1rw5ffdqcq"></a>Phusion

<a name="h.dczrad3ll1mv"></a>10

<a name="h.kzxwyaz9wz9"></a>10

<a name="h.2vqbu3sc5ab"></a>10

<a name="h.wu32cplrcglv"></a>10

<a name="h.wirojc6d4snp"></a>10

<a name="h.pscsdxa33omg"></a>10

<a name="h.z6vdw2jwwuqx"></a>10

<a name="h.pnt4xdtz05ne"></a>10

<a name="h.123cxn5gbnrc"></a>10

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

<a name="h.ovdj40su5h7d"></a>Water

<a name="h.ttmmg3mlbr01"></a>7

<a name="h.7yuio2gunhyy"></a>7

<a name="h.lfps3rbohioe"></a>7

<a name="h.vvbnf0qym3r"></a>7

<a name="h.eu2pv3427mvz"></a>7

<a name="h.c0yscubylyim"></a>7

<a name="h.ais0f5cxkimo"></a>7

<a name="h.ijkk50bc8vfc"></a>7

<a name="h.1z3sb2ooz031"></a>7

<a name="h.c9f2uuw3g1k9"></a>7

<a name="h.f9vr6a4tau82"></a>

<a name="h.2v8293oxl31m"></a>7/27/2013

<a name="h.3yl3n1uiaccl"></a>

<a href="#" name="104372c6427563de636b62fe345d95e734a96dae"></a><a href="#" name="8"></a><tbody></tbody>

<a name="h.lif5uh8a9ru5"></a>1

<a name="h.skiksmnn8a6s"></a>i5-13-1

<a name="h.z7jenu3w39sb"></a>18.4

<a name="h.z0hca4r5b8qn"></a>2

<a name="h.f3nxtwm21n6m"></a>i5-13-2

<a name="h.uqvy6aqkj78d"></a>9.7

<a name="h.axhxqpai8z1f"></a>3

<a name="h.rhsnpzxmd1h8"></a>Failed

<a name="h.9vgxdsujxymk"></a>---

<a name="h.e1oongaoz4hm"></a>4

<a name="h.44vk83d784r1"></a>i5-13-3

<a name="h.m7xvhd894v72"></a>6.6

<a name="h.2l8b57cblu"></a>5

<a name="h.ahsuxtk55dkw"></a>i5-13-4

<a name="h.ncz5e124gpbc"></a>3.0

<a name="h.od8av1xynej2"></a>6

<a name="h.c96gtq9s71y2"></a>i5-13-5

<a name="h.vxhbmifkq0pu"></a>4.7

<a name="h.6p6boqiu7y13"></a>7

<a name="h.5ncuhw1ecuin"></a>i5-13-6

<a name="h.wkv6va6u5opu"></a>6.2

<a name="h.za7l81wf53cb"></a>8

<a name="h.gvxjhuaekgc5"></a>Failed

<a name="h.m2gww3x6td12"></a>---

<a name="h.ujpyzzvmoq1h"></a>9

<a name="h.tfh4pztzb4ta"></a>i5-13-7

<a name="h.c0czudjusars"></a>6.2

<a name="h.3z9hssn7h1v1"></a>10

<a name="h.jtgwcyr3oldj"></a>i5-13-8

<a name="h.mavxc2ewd4cr"></a>7.3

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

<a name="h.hbpr7r9p3d9"></a>

Retrieved from "http://2013.igem.org/Team:UT_Dallas/week4"