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Team:Technion/20 August 2012
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==Inbal== | ==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.<br> | ||
| + | - 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.<br> | ||
| + | |||
==Asaf== | ==Asaf== | ||
Revision as of 21:09, 22 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
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
Shahar, Evgeni & Inbal
- PCR of all Chris RNAPs with pPROLAR and pSB1A2/C3 suitable primers - Checking all PCR products on agarose gel 2%
