Dear everyone:
I am trying to denature dsDNA plasmid drived from pUC19. When I heated
it using PCR machine at 94C for 5 min., immediately transfered into ice
water, and tried agarose gel electrophoresis, it showed smear band.
When I had tried SSCP with FastSystem(Pharmacia), I once had read in the
manual that the mobilities of denatured ssDNA are different depending on
the electrophoresis temperature(the system can retain the temperature of
gel running). So, I guess the smear band is because of the temperature
increment during electrophoresis to cause variant changes of single
strand conformation(my electrophoresis kit cannot regulate temperature).
How can I get ssDNA band by agarose gel electrophoresis? Any reply would
be appreciated. Thanks.
Sincerely,
Park, Seung Kyu
mailto:parksk at kistmail.kist.re.kr
Biomedical Research Center, KIST, 39-1,
Hawalgok-dong, Sungbuk-gu, Seoul, 136-791, Korea
Tel : +82-2-958-5907 Fax : +82-2-958-5909 Beeper : 015-8516-2512
