Search found 7 matches

by antonyviridae
Mon Sep 12, 2011 9:25 pm
Forum: Molecular Biology
Topic: DNA methylation
Replies: 2
Views: 9860

In regards to the prokaryotic methylation, it also work in reverse. When DNA is methylated, some restriction enzymes are unable to cut the DNA. What do you mean by how to compare them? Their both secondary modifications to DNA, for one.
by antonyviridae
Wed Sep 07, 2011 8:33 pm
Forum: Molecular Biology
Topic: genomic rescue construct from BAC DNA
Replies: 2
Views: 5505

Re: genomic rescue construct from BAC DNA

If I were you, I wouldn't just add more DNA to the reaction. I've come to believe that if you can see it, you can clone it. Are you using the right buffer for your restriction enzymes? For Fermentas enzymes, they suggest 2x tango but the BamHI is reduced to 50-100% activity. What company are the enz...
by antonyviridae
Wed Sep 07, 2011 2:32 pm
Forum: Molecular Biology
Topic: Lipofectin/Labfectin lethality towards insect cells
Replies: 5
Views: 7813

Re:

what about membrane disruption? That's what most people think about it. So in the reaction, the phosphatidylethanolamine (dope) would bind to the plasma membrane and fuse to bring in the DNA, but how does it cause lethality? Does it destablize the membrane while fusing? Another source claims it's t...
by antonyviridae
Wed Sep 07, 2011 2:13 pm
Forum: Molecular Biology
Topic: PLease Please I need help with my PCR! Greatly appreciated!
Replies: 8
Views: 13134

Re:

increasing the volume of reaction? I thought, that it's better to have smaller volume, since the temperature changes are faster and more accurate. Though it may be true that a smaller volume would help with temperature changes, increasing the reaction volume also increases the amount of reagents th...
by antonyviridae
Tue Sep 06, 2011 4:31 pm
Forum: Molecular Biology
Topic: PLease Please I need help with my PCR! Greatly appreciated!
Replies: 8
Views: 13134

Re: PLease Please I need help with my PCR! Greatly appreciated!

From what I can see, your reaction looks good. Some suggestions that I have are: -Don't use a gradient for the annealing. One temperature two-four degrees lower than your lowest primer Tm will suffice. Doing this might also get rid of those non specific bands. -You need at least two-four nucleotides...
by antonyviridae
Fri Sep 02, 2011 4:17 pm
Forum: Molecular Biology
Topic: Lipofectin/Labfectin lethality towards insect cells
Replies: 5
Views: 7813

Lipofectin/Labfectin lethality towards insect cells

What is it about Lipofectin/Labfectin that makes them so lethal for insect cells during transfection? -L-alpha-phosphatidylethanolamine, dioleoyl -Dimethyldioctadecylammonium Bromide I understand that we are using a lipid and a molecule that can act as a lipid but what do they do to kill the cells s...
by antonyviridae
Fri Sep 02, 2011 4:06 pm
Forum: Molecular Biology
Topic: PCR and Gel conundrum
Replies: 3
Views: 2999

PCR and Gel conundrum

Why is it that when I run the exact same PCR reaction (with the same sample) on two different PCR platforms, that when I run it on a gel or on Qiaxcel I see that the bands are running at different sizes? The difference isn't significant but enough to be noticable.