Improved electrophoresis of nucleic acids. - (Aug/09/2008 )
QUOTE (swanny @ Aug 14 2008, 02:53 AM)
The borate system works very well for short, fast gel runs, but I wouldn't recommend it if you have a large gel format and you want to run for a long time.
I run 15cm gels at 150V for 5 hours for my work. With SB the gel completely melted away into the buffer!!
Went back to TBE, no problems at all.
SB is not good for long runs!
-MTO-
QUOTE (JCbio @ Aug 19 2008, 10:38 AM)
Does anyone know if using the SB buffer would affect a subcloning procedure? We use TAE now and it works fine, but it would be great to run it faster if at all possible. I just don't want to cut the gel out and have the SB mess up the finicky nature of DNA Ligase or disturb the balance of the ligation buffer.
-Jerry
-Jerry
I tried the SB buffer during the cloning of a few vectors. It was perfect.
-scolix-
We also tried SB for electrophoresis in our lab. We run 1% gel at approx 300V for about 15 minutes. The result is amazing. It gave the same results as running in TAE buffer at much lower voltage!
I prepared a 25x stock solution (125mM). The sodium borate does not dissolve very good.. But after warmimg up it was ok!
Regards
-JacksonCreil-
QUOTE (JacksonCreil @ Aug 22 2008, 10:29 PM)
We also tried SB for electrophoresis in our lab. We run 1% gel at approx 300V for about 15 minutes. The result is amazing. It gave the same results as running in TAE buffer at much lower voltage!
I prepared a 25x stock solution (125mM). The sodium borate does not dissolve very good.. But after warmimg up it was ok!
Regards
I prepared a 25x stock solution (125mM). The sodium borate does not dissolve very good.. But after warmimg up it was ok!
Regards
Hello.
I run SB for electrophoresis using 0.8% gel at 250V for 10 min but the band separation is not good. I increased the time to 15 minutes but the band became not intense with some smearing. The ladder i was using is 100 bp DNA ladder plus from Fermentas.
Any suggestion to solve my problem? Thanks.
-dcch-
An 0.8% gel will not resolve 100 bp ladders with any buffer. You need a 1.5% to 2% gel for short fragments.
-phage434-
QUOTE (phage434 @ Aug 24 2008, 11:32 AM)
An 0.8% gel will not resolve 100 bp ladders with any buffer. You need a 1.5% to 2% gel for short fragments.
Sorry, phage, but I have to disagree with you there. The SB system I use is 0.8 - 0.9%, and I resolve the members of the 25 bp ladder. I must add, though, that the separation between 200 and 300 bp gets a bit iffy, unless you run the gel all of the way out.
-swanny-