Two Sequencing questions:
robert at arbo.microbiol.uwa.oz.au
Wed Aug 12 22:44:18 EST 1992
In article <zsha.713469334 at sanger.bb.iastate.edu> zsha at iastate.edu (Zhengyu Sha) writes:
>In <1992Aug10.154723.23852 at magnus.acs.ohio-state.edu> rrumpf at magnus.acs.ohio-state.edu (Robert Rumpf) writes:
>>I have two questions regarding sequencing protocols:
>>1. Our 35-S gels are still sticky after drying; the saran wrap sticks to them
>>and is difficult to remove. Why? We also noticed that removing the saran wrap
>>and placing film directly on the gels apparently causes some damage to the
>>film from the moisture or urea in the gel...
This is the nth time I've heard of people wanting to remove the saran wrap
of gels containing 35S. Why ? We don't fix our gels, nor take of the wrap. We
peel the gels of the glass with Whatman 3MM and dry them under wrap in a vacuum gel dryer. Than expose them to Fuji RX (not a very sensitive film !) and get good results from an overnight exposure. Just make sure your reactions are good (CLEAN template and good template to primer ratio)
>>2. Anyone have any experience with 33-P yet? We would like to know if the
>>energy from this isotope is sufficient to pass through saran wrap so we can
>>expose films without taking the wrap off.
> You mean 32-P? The answer is yes. Even with 35-S sequencing gel we exposure the dried gel without removing the saran wrap. The results are satisfied if you labeling reaction is good.
We are about to develop some film from a 33P run. We'll let you know tomorrow
what our findings are.............................
Robert (semi-mature Gouda CHEESE) Coelen
Dept of Microbiology
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