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Smearing of the PCR Products - (Jun/07/2006 )

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check your amplicon - length AT-GC secondury structures (hairpins)
GC-rich - betaine AT- rich - magnesium
ELONGATION time -- 25 seconds for 1 kb for Taq ( do not ovrhold at any!!!!!!! temperatures)

Try ammonium PCR buffer or use GoTaq Green flexy(promega) - it is robost and
resist wide range of magnesium concentrations
if DNA is DAMAGED - Taq +Pfu ( Long PCR blend ) helps

Touchdown - Stepdown - Hotstart will help

Use cycler with ramping rates 3 C / sec or more under "tube control" heated lid
reaction volume - 20 mkl- PCR plastics from Abgene


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