Team:TUDelft/28 July 2009

From 2009.igem.org

Lab Notebook

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28 July 2009

Sriram

Laura said that the RbCl needed for the new protocol for competent cells preparation is out of stock we cant prepare chemically competent cells till we get it (earliest possible on thursday). Also it seems we can get the plasmid backbones amplified and purified only by thursday morning it is fine. But if we dont get the RbCl on thursday we planned to do electroporation for the assembly done on that day. We made a plan to prepare the electro competent cells tomorrow and perform a test electroporation by tomorrow itself. Discussed with Laura about the use of the BioTek Synergy instrument which will be used for making measurements of the delay device working and she gave consent to help us by giving a demo of its use in next week by performing a test assay (GFP expression with growth). She too doesn't know too much about the instrument, which is the reason for this test assay.

Daniel

Performed transformation of the 6x25 µl ccdB resistant competent cells with the 6 plasmid backbones that will be used in the project and plated with corresponding antibiotics.

Calin

Finished gel from yesterday. Looked at protocol for making cells electrocompetent. Miniprep to extract pSB1AK3.

Gel28072009-sd-delay-conj.png


Well Part Expected Plasmid Size Status
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10 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
11 BBa_S03473 λp-RFP
12 BBa_K081013 RBS-cI-RBS 2898
13 BBa_J23008 key3c 2450
14 BBa_B0015 Double Terminator 3318
15 BBa_S03335 λp-GFP
16 BBa_R0040 pTet 2133
17 BBa_J23031 lock3c 2398
18 BBa_E0840 GFP generator 2957
19 BBa_J23100 strong promoter 2948
20 BBa_I13522 OriT-R 2357

Tim Vos

Extracted the plasmid DNA for the cultures incubated yesterday for 3 bio-bricks λp-GFP [R0040], RBS[B0034] and pLacI[R0010].