Search found 6 matches

by patlangic
Mon Aug 08, 2011 8:26 pm
Forum: Molecular Biology
Topic: Analyzing gel electrophoresis picture?!
Replies: 4
Views: 10444

one correction to the previous message:

''...that particular cut site is mutated at both loci..''

i meant 'alleles' of course; presence of different loci have nothing to do with this case :)
by patlangic
Mon Aug 08, 2011 8:19 pm
Forum: Molecular Biology
Topic: Analyzing gel electrophoresis picture?!
Replies: 4
Views: 10444

Re: Analyzing gel electrophoresis picture?!

it looks like a picture representing RFLP analysis of a 250bp-long DNA fragment on which one polymorphic restriction cut site is located (at around 175th bp). these three samples should have been collected from three different individuals. lane2 and lane3 represents the two homozygous genotypes (let...
by patlangic
Thu Apr 07, 2011 5:36 pm
Forum: Molecular Biology
Topic: pET28a(+) frustrations
Replies: 1
Views: 4028

Are you sure that the Shrimp Alkaline Phosphatase you used is really working? And how did you recover (or purify) your plasmid after dephosphorylation? Have you tried ethanol precipitation or something? I also use pET28a(+) as the expression vector but dephosphorylate it by rAPid alkaline phosphatas...
by patlangic
Tue Dec 21, 2010 2:21 pm
Forum: Molecular Biology
Topic: optimum molar ratio for pET28 cloning?
Replies: 5
Views: 2528

Re: optimum molar ratio for pET28 cloning?

Kezzer wrote:Hey, I used PET28c using a molar ration of 1:8 if 1:3 isn't suitable maybe try that :)


what a coincidence :) I just tried 1:7 and 1:3 ratios and performed transformation, so I'll get the results in a couple of days. Thank you..
by patlangic
Tue Dec 07, 2010 12:24 pm
Forum: Molecular Biology
Topic: optimum molar ratio for pET28 cloning?
Replies: 5
Views: 2528

OK, I'll try it. Thank you :)
by patlangic
Tue Dec 07, 2010 9:47 am
Forum: Molecular Biology
Topic: optimum molar ratio for pET28 cloning?
Replies: 5
Views: 2528

optimum molar ratio for pET28 cloning?

Hello everyone..I wonder if someone could help me find the optimum molar insert:vector ratio (or ratio range) to clone my gene into the pET28a(+) vector. If there is anyone who used this expression vector before, I would appreciate any recommendation. Thanks in advance..