Team:TUDelft/31 July 2009

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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

Today I tested the competent cells prepared yesterday. For that I chose a three DNA samples. One is a diluted biobrick λp-rep for testing electro-competent cells, λp-R-GFP-T which had very high DNA concentration and biobrick pSB1K3 containing mRFP which had a normal DNA concentration for the two chemically competent cells prepared by TSS and TMF(RbCl) buffers respectively.

Orr helped me in doing the transformation of chemical competent cells prepared by TSS buffer. I did the transformation of those prepared by TMF buffer and electroporation. I have kept the plates in the draw to check the results on monday.

Calin

R751 cells arrived today. 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

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