"
Team:Calgary/11 August 2010
From 2010.igem.org
Raidakhwaja (Talk | contribs) |
|||
| Line 5: | Line 5: | ||
Today I did a miniprep of the pRFP colonies because I ran out. Then I biobricked the pRFP using a gradient PCR with an annealing temperature of 55 degrees to 73 with 8 tubes in a 12 well gradient placed evenly. The PCRs that are run include 1.0 mM of MgCl2, 1.5 mM of MgCl2 (regular), 2.0 mM of MgCl2 using the new biobrick primers with NotI and XbaI forward sites and SpeI and NotI reverse sites. The tube of pRFP used was from colony 3 (C3). I also ran a 1.3 mM MgCl2 with the old primers with EcoRI, NotI, XbaI and the SpeI on the reverse. I also did a colony PCR of the pRFP plasmid switch (old PCR purified product and old primers). I ran the gel and the results are..... I also did my article for the blog. | Today I did a miniprep of the pRFP colonies because I ran out. Then I biobricked the pRFP using a gradient PCR with an annealing temperature of 55 degrees to 73 with 8 tubes in a 12 well gradient placed evenly. The PCRs that are run include 1.0 mM of MgCl2, 1.5 mM of MgCl2 (regular), 2.0 mM of MgCl2 using the new biobrick primers with NotI and XbaI forward sites and SpeI and NotI reverse sites. The tube of pRFP used was from colony 3 (C3). I also ran a 1.3 mM MgCl2 with the old primers with EcoRI, NotI, XbaI and the SpeI on the reverse. I also did a colony PCR of the pRFP plasmid switch (old PCR purified product and old primers). I ran the gel and the results are..... I also did my article for the blog. | ||
| + | |||
| + | <u> Raida </u> | ||
| + | Today, I ran a 1% agarose gel electrophoresis of my PCR from yesterday. The PCR was done with Ssdeleted forward primer and reverse primer on malE31 template and also malE31 without the signal sequence and MalE as a positive control. The results were positive for malE31delSS and also MalE. So Emily PCR purified the PCR product. The next step will be doing restriction digests and also cloning into a TOPO vector. | ||
}} | }} | ||
Revision as of 21:39, 11 August 2010
Wednesday August 11, 2010
Jeremy
Today I did a miniprep of the pRFP colonies because I ran out. Then I biobricked the pRFP using a gradient PCR with an annealing temperature of 55 degrees to 73 with 8 tubes in a 12 well gradient placed evenly. The PCRs that are run include 1.0 mM of MgCl2, 1.5 mM of MgCl2 (regular), 2.0 mM of MgCl2 using the new biobrick primers with NotI and XbaI forward sites and SpeI and NotI reverse sites. The tube of pRFP used was from colony 3 (C3). I also ran a 1.3 mM MgCl2 with the old primers with EcoRI, NotI, XbaI and the SpeI on the reverse. I also did a colony PCR of the pRFP plasmid switch (old PCR purified product and old primers). I ran the gel and the results are..... I also did my article for the blog.
Raida Today, I ran a 1% agarose gel electrophoresis of my PCR from yesterday. The PCR was done with Ssdeleted forward primer and reverse primer on malE31 template and also malE31 without the signal sequence and MalE as a positive control. The results were positive for malE31delSS and also MalE. So Emily PCR purified the PCR product. The next step will be doing restriction digests and also cloning into a TOPO vector.
