cleaning up PCR reactions
jgraham at bronze.ucs.indiana.edu
Thu Jul 1 10:13:43 EST 1993
.....cleaning up PCR reactions.......
I can demonstrate the removal of all detectable P32 end labeled primers from
500 bp PCR fragmnets by a simple single step isopropanol precipitation. Ed and
I are using 1/5 vol of 10 M NH4Oac and 1 vol of isopopropanol. Spin
immediately at room temperature for 20 minutes. If I am going to use this
for sensitive (to NH4 ?) E. coli RNAP in vitro transcriptions, I pass them
through a G-50 spin column as described in "the Maniatis". Another lab here
uses templates prepared this way for direct sequencing with alternate primers.
Why pay more ?
Jim "kit-free" Graham
Biology and Chemistry Departments
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