Search found 4 matches

by Duke
Tue Oct 02, 2007 11:37 am
Forum: Molecular Biology
Topic: Please explain to me - CMVp
Replies: 4
Views: 6113

Hi. Thinking about this T7 and CMV promoter again... I have both in my plasmid. I have understand that T7 polymeras can be used for in vitro/cell-free trancription. Is that what it is used for? And CMV promoter as I understood it is good because than you dont need to make a RNA transcript of your g...
by Duke
Tue Oct 02, 2007 9:32 am
Forum: Molecular Biology
Topic: cloning problems
Replies: 7
Views: 4367

Re: cloning problems

hi! I´m trying to clone different 1,5 kb dna fragments into the pGemT-Vector and in a pQE-Vector. One time the cloning into the pGem worked. But after this success no ligation worked any more. I don´t know what is the problem. For cloning into the pGem I use a DNA polymerase that makes A overhangs....
by Duke
Tue Oct 02, 2007 9:11 am
Forum: Molecular Biology
Topic: Enzymes Question
Replies: 8
Views: 9773

Re: Enzymes Question

What two things can cause enzymes to malfunction? I couldn't find the answer in my texbook. If malfunction means not the function which it should have and not loss of function, then I have 2 examples for you: Solvent can inlfuence the substrate specificity of some enzymes (Restriction enzymes somet...
by Duke
Tue Oct 02, 2007 7:56 am
Forum: Molecular Biology
Topic: priciple of cloning
Replies: 5
Views: 5069

Re: priciple of cloning

Hi I´m trying to create a clone and have some ligation problems. And while I am thinking about the reasons for these problems and looking on my gels- I have one question. When I run a gel with my ligation-mix there is a lot of smear. And it seems to be logical because my cut insert can ligate with ...