An IPCR Tale
SPAGNOL at PLAUTO.CSATA.IT
SPAGNOL at PLAUTO.CSATA.IT
Sat Mar 11 09:57:04 EST 1995
Dear colleagues,
here is the story of my IPCR:
First try: (1.5 mM MgCl2): smear
Second try: (MgCl2 optimized to 4.5 mM): smear, plus
very,very faint band at 600 bp.
Third try: ( Annealing T brought from 52 C to 60 C):
smear, plus faint band at 600 bp
Fourth try: ( using the "stick in the toothpick" method
to isolate the 600 bp band ): VERY CLEAR band at 600 bp. Still the
smear, even if much fainter, is still there.
Question: should I ignore the smear, go ahead and subclone the
product, or try to get rid of it? And how?.Thanks.Giorgio.
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