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Sticky Problem to a Blunt end


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#1 Kausik Chakrabarti

Kausik Chakrabarti

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Posted 12 August 2002 - 11:19 PM

I have an insert which has got at one end a blunt end restriction site & on the other hand a sticky end restriction site. After PCR amplification of the insert, I am not able to get Proper Fall Out  after restriction digestion & therefore no colonies after subsequent cloning & Ligation. Please Help

#2 Wotan

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Posted 13 August 2002 - 08:00 AM

Where are the restriction sites inthe PCR-product?
If they are in the primers, have you added enough bases at both sides of the restriction sites?




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