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help in long pcr - (Feb/24/2014 )

Are there any tips on amplifying >5kb fragment?

-Angeline-

Use a new enzyme (Phusion or Q5). Use specific primers. You may need additives for high GC DNA (5% of a 1 M betaine solution works well).

Make sure your extension times are long enough (easy with these enzymes).

-phage434-