Team:TUDelft/31 July 2009

From 2009.igem.org

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(Sriram)
(Sriram)
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===Sriram===
===Sriram===
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I planned to test the competent cells today. For that I chose a three DNA samples. One is a diluted biobrick λp-rep for testing electro-competent cells, λp-GFP which had very high DNA concentration and cI which had a normal DNA concentration.
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I planned to test the competent cells today. For that I chose a three DNA samples. One is a diluted biobrick λp-rep for testing electro-competent cells, λp-GFP which had very high DNA concentration and biobrick pSB1K3 which had a normal DNA concentration.
===Calin===
===Calin===

Revision as of 22:00, 3 September 2009

Lab Notebook

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31 July 2009

Tim Weenink

Did some minipreps today. Then nanodropped them:

Biobrick name Internal reference Concentration ng/µl 260/280 260/230
BBa_K142205 *S2 86.4 2.01 2.03
BBa_K142205 *S3 107.7 2.03 2.11
BBa_K142205 *S4 69.0 1.97 1.96
BBa_K142205 *S5 87.4 1.98 1.99
BBa_K142202 *T1 128.5 1.99 2.09
BBa_K142202 *T2 129.4 2.02 2.16
BBa_K142202 *T3 126.6 1.97 2.06
BBa_K142202 *T4 106.7 1.95 2.05
N/A  !A 171.2 1.98 2.11

Sriram

I planned to test the competent cells today. For that I chose a three DNA samples. One is a diluted biobrick λp-rep for testing electro-competent cells, λp-GFP which had very high DNA concentration and biobrick pSB1K3 which had a normal DNA concentration.

Calin

Cultures in all pSB4C5 tubes. Did miniprep on all tubes and also made a plate.

Part Concentration ng/µl 260/280 260/230
pSB4C5 I 123.0 1.91 2.06
pSB4C5 II 125.8 1.92 2.29
pSB4C5 III 130.7 1.95 2.07

Todays gel confirmed that oriT-R digestion failed and pSB4C5 is way too large.

Gel31072009.png


Well Part Expected Plasmid Size Status
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22 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
23 oriT E + P digest 2079
24 pSB4C5 I 3896
25 pSB4C5 II 3896
26 pSB4C5 III 3896

R751 cells arrived. Made two plates without antibiotics.

Digestions were started:

position parts volume in µl
Upstream BBa_I13522 pTet-GFP 27.0
H2O 15.5
Upstream BBa_E0840 rbs-GFP-term 5.0
H2O 37.5
Backbone pSB1C3 5.0
H2O 37.5