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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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Nonspecific insert in TOPO TA cloning
Posted by: egugene (IP Hidden, New member, 1)
Date: July 13, 2005 08:12AM
I tried to clone part of human gene using TOPO TA kit from invitrogen. First desired band of PCR product from gel was extracted and purified. Then it was cloned using TOPO TA kit. Screening of colonies revealed bands of expected size. But plasmid sequencing showed that between gene-specific primers there is a sequence which doesn't have common with cloned gene. Can You help and suggest where is the botlle neck of this problem, what can be the reasons and how to solve it. Thank You beforehand
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