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Published online Jan 24, 2006. doi:  10.1186/1472-6750-6-7

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Design for producing wild-type and mutant shRNA vectors simultaneously. A forward strand of the wild-type hairpin (blue) is synthesized together with a reverse strand containing a one bp mutation within both the sense and antisense copy of the target sequence (shown in red). The double stranded hybrid is ligated into the retroviral vector 5' of an H1 promoter and transformed into competent bacteria. Since replication is semi-conservative, the daughter bacteria will be of two different populations that carry either a double-stranded wild-type or a double-stranded mutant vector and can be isolated by preparing and sequencing individual colonies.

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