Cyclic Sequencing

mmelan at mmelan at
Fri Jan 29 18:14:34 EST 1993

        With regard to the question of cycle sequencing......

        I have never used Stratagene's kit but the lab next door has NEVER
gotten it to work particularly with internal incorporation of isotope.
        I have had good luck with Gibco's kit using 32P end labeling.  The
sequence obtained is clear (not big fat fuzzy bands) after overnight exposure
at room temp with no screen.  You should, however, use high specific activity
gamma 32P.....lower activity label gives pale sequence.

Melissa Melan                                   mmelan at
Department of Biological Sciences
Wellesley College
Wellesley, MA  02181
I have no connections with Gibco or Stratagene......only opinions.

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