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Team:Technion/20 August 2012
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Ilya
- Minipreped R0062+Fus1 in pSB1C3 and in pSB3C5, Fus2 in pSB1C3 and in pSB3C5, Fus1 in pSB1C3. Forgot to do a glycerol stock.
- Diluted the new primers.
- PCR to delete the riboswitch from R0062+Fus1 in pSB1C3 and in pSB3C5, Fus2 in pSB1C3 and in pSB3C5.
- Checked PCR products on a gel. Got the correct bands in all 4 reactions.
- Cleaned the PCR products using the new kit with the tips. Got good concentration but poor 260/280 and 260/230 ratios.
Inbal
-The primers for chris's RNAP are finaly arrived! So I diluted the primers for the T7* RNAP gene: the primers corresponding to the plasmid pSB1C3, and primers corresponding to the plasmid pPROLAR. The primers were designed by Rachel, Evgeni, Shachar and me.
- PCR to T7* gene using the primers above. I expected a product size of approximately 2700bp.
- Running the products on a 1% agarose gel, the results: not successful,there wasn't a band nearly 2700bp, in any of the primers mentioned above.
Asaf
Doing a PCR reaction for extracting the RiboSwitch from the plasmid with different overhangs.
Hila
- hybridization for oligos XbaI_MCS_suffix_A+AS.
- restriction reaction with XbaI + PstI (HF) to plasmid pSB1C3 and the hybridization products.
- 2% gel for hybridized and cut oligos.
- 1% gel for plasmid backbone.
the gel for the plasmid does not look good.
- starter for the glycerol stoke of pSB1C3.
tomorrow I will miniprep and restrict the plasmid again.
Lior
Noa
PCR purification for ALP and pT7.
Cutting plasmid pT7 and Inserts (ALP and xyIE) with XmaI and XhoI.
Ligation of pT7 with ALP and xyIE
.
Shahar, Evgeni & Inbal
- PCR of all Chris RNAPs with pPROLAR and pSB1A2/C3 suitable primers - Checking all PCR products on agarose gel 2%
