HELP! Odd Gel Pattern!!
smori at opus.nmsu.edu
Fri Jun 3 01:32:14 EST 1994
brunker at delphi.com wrote:
: Instead of getting descrete bands on our electrophoresed agarose gel, we've
: been getting "smeared" patterns...a dark band followed by a gradient of lighter
: stains trailing back to the well. (I hope I'm describing this clearly).
: Does anyone know what causes this, and how we fix it?
: These are PCR products digested by Nco1, and should produce not more than
: three nice strips each, not counting the primer lines. The marker lane looks
: fine. The pieces we're looking at (we hope) are ~30, ~190 and `160 pb's.
: Has this happened to anyone else? Are we really producing an increasingly
: large range of DNA strands?
: How could such a fate have befallen us?!!
: Marla Brunker
: brunker at delphi.com
What maybe wrong is that your PCR was not specific enough and you have
gotten some non-specific amplified DNA.
Is your PCR from Genomic DNA? Do you have highly specific primers?
NCO1 may be cutting the contaminant amplified elements, thereby giving you
smearing. I know this sounds too weird but who knows.
Program in Molecular Biology
Dept. of Chemistry and Biochemistry
NMSU Las Cruces NM
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