PCR cloning - how many clones must one sequence?

Bo Johansen boj at bot.ku.dk
Thu Mar 25 05:18:34 EST 2004


We are cloning products from a nested PCR and single base variation 
occur in many clones. We use degenerated primers and taq polymerase for 
both PCR reaction. How many "alike" clones do we have to sequence in 
order to be sure that we have the correct sequence?
Will it help if we use a proofreading enzyme instead of taq?

Bo





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