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SB Buffer and Page Electrophoresis


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#1 stefan_k

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Posted 22 September 2010 - 01:15 AM

I was wondering if somebody could help me.
I use SB Buffer for my Agarose Gel electrophoresis.
Due to the good results i would like to use SB buffer for Page electrophoresis as well.

Are there any pitfalls with SB buffer and page electrophoresis.

Is it possible to use the same loading dye for SB buffer and TBE buffer.

Do I have to mind something else.

Thank you guys,

Stefan

#2 mdfenko

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Posted 22 September 2010 - 06:16 AM

you should be able to substitute sb for tbe in page. try it first with less precious samples. take care to follow similar parameters as with tbe (eg-temperature).

if you are using the same loading dye in age with sb or tbe then you should be able to do the same with page.
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#3 stefan_k

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Posted 27 September 2010 - 07:50 AM

you should be able to substitute sb for tbe in page. try it first with less precious samples. take care to follow similar parameters as with tbe (eg-temperature).

if you are using the same loading dye in age with sb or tbe then you should be able to do the same with page.


Thanks alot for the immediate reply.

I tried the SB puffer today and didn't get good results.

I have attached a picture of the gel.

Does anybody have an idea?

sb_test.jpg

#4 tfitzwater

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Posted 27 September 2010 - 08:56 AM

I compared 8% acrylamide/8 M urea gels using 1x TBE or 1x SB in 2004. Samples included a concentration gradient of a 25 mer oligo and the Bio-Rad AmpliSize DNA ladder (50 - 1000 bp) denatured and loaded in a typical formamide dye sample buffer. Gels were run at 60 watts for 28 and 27 minutes, stained with SYBR Gold, and imaged on a Fuji phosphorimager. Buffer temperature started at 25C and increased to 49C for TBE and 52C for SB. XC traveled 6.5 cm and 7.7 cm, respectively. The 25 mer traveled 12.8 cm and 13.2 cm, respectively. BPB traveled 14 cm and 14.7 cm, respectively. Resolution was identical except for the spacing on the 600 - 1000 bp bands in the DNA ladder. I did not see any artifacts or distorted bands.




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