Search found 23 matches

by curious1
Tue Oct 02, 2007 4:51 pm
Forum: Molecular Biology
Topic: Can siRNA's be thawed on 37 heat block?
Replies: 0
Views: 991

Can siRNA's be thawed on 37 heat block?

Just curious, can siRNA's be thawed at 37? Since total RNA's always have to be thawed on ice, does the same apply for siRNA since it's still RNA?
by curious1
Wed May 30, 2007 6:15 pm
Forum: Molecular Biology
Topic: incomplete transfer of protein samples for western blot
Replies: 6
Views: 11885

hi blcr11, Thanks for your reply! hmmm, the blot looks OK, the ladder transfers over just fine but since I am trying to detect a very small MW protein (~18kd) I do worry about the proteins tranferring right through. Standard protocols tell me to run at 100V which is what I do, but the current is alw...
by curious1
Thu May 24, 2007 9:22 pm
Forum: Molecular Biology
Topic: incomplete transfer of protein samples for western blot
Replies: 6
Views: 11885

Ah, it turns out it was a loose wire. I've connected the wire back and it's running, but the mAmps seem a bit high, around 400mAmps. Is that going to be a problem?
by curious1
Thu May 24, 2007 3:16 pm
Forum: Molecular Biology
Topic: incomplete transfer of protein samples for western blot
Replies: 6
Views: 11885

incomplete transfer of protein samples for western blot

Hi Everyone, I recently starting having trouble getting complete transfers on my western blots. I transfer at 1 hour at room temperature at 100V and it has always worked fine. I made fresh transfer buffer and tried transferring overnight, even used a new power source and still the proteins don't tra...
by curious1
Thu Feb 01, 2007 9:58 pm
Forum: Molecular Biology
Topic: shRNA versus siRNA
Replies: 4
Views: 17883

o i understand now, thanks!
by curious1
Wed Jan 31, 2007 4:25 pm
Forum: Molecular Biology
Topic: shRNA versus siRNA
Replies: 4
Views: 17883

O I see, Does that mean shRNA is transcribed along with the genome while siRNA attacks the after double stranded RNA's have already been transcribed. Also, is it true that siRNA can only be used for transient transfections while shRNA for stable transfections since siRNA is not incorporated into the...
by curious1
Wed Jan 31, 2007 4:56 am
Forum: Molecular Biology
Topic: shRNA versus siRNA
Replies: 4
Views: 17883

shRNA versus siRNA

Hi hi, I was wondering what the difference between using shRNA and siRNA really is. I have they both use the same mechanism for silencing except that shRNA contains an extra hairpin structure, but is that all? What is the advantage of using shRNA rather than siRNA? I read that shRNA is better for st...
by curious1
Tue Jan 30, 2007 6:29 pm
Forum: Cell Biology
Topic: How do you make a drug-resistant cell line? determine IC50?
Replies: 0
Views: 2534

How do you make a drug-resistant cell line? determine IC50?

Hi all, I am trying to study for chemotherapy resistance for two drugs. I was wondering if anyone knew the exact protocol for developing a drug resistant cell line. From the articles that I read they only say that drug resistant cell lines were produced from stepwise additions of the drug of interes...
by curious1
Fri Sep 29, 2006 3:12 pm
Forum: Molecular Biology
Topic: What antibody to use? Conjugated vs. unconjugated?
Replies: 0
Views: 2385

What antibody to use? Conjugated vs. unconjugated?

Hi, Sorry if this seems to be a simple question but how do we know what kind of antibodies to use? I mean what is the difference between conjugated and unconjugated and how do we know which one to use? So, if we have an unconjugated primary antibody do we need to use an unconjugated secondary? Also,...
by curious1
Thu Sep 28, 2006 5:13 pm
Forum: Molecular Biology
Topic: protein samples running in a straight line
Replies: 3
Views: 1896

Hi, I used a 15% gel so the acrylamide concentration should be sufficient, this is the first time I'm doing a western blot and no matter how many times I read the protocol I just seem to have a million questions and uncertainties. Seems like nothing is going the way it's supposed to. So I stained th...
by curious1
Tue Sep 26, 2006 6:14 pm
Forum: Molecular Biology
Topic: protein samples running in a straight line
Replies: 3
Views: 1896

protein samples running in a straight line

Hi everyone,

I am recently running SDS gels with protein samples for western blots. However, I noticed that my gels have been running strangely, that is the protein samples are all running in a straight line! Has anyone ever experienced this before? What could this be from? Please advise!
by curious1
Tue Aug 22, 2006 11:02 pm
Forum: Molecular Biology
Topic: Horse serum vs. Fetal Bovine Serum in DMEM?
Replies: 1
Views: 3086

Horse serum vs. Fetal Bovine Serum in DMEM?

Hi everyone, I was wondering if anyone knew whether it is ok to use horse serum in place of fetal bovine serum when preparing DMEM media. We usually add FBS but we ran out so I had added about 2ml of FBS and then used 23ml of horse serum to make my media. Also, is it okay to mix the two serums? Than...