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Plasmids Sequence And Ligation Independent Cloning

This post categorized under Vector and posted on June 10th, 2019.
Gene Vector PCR: Plasmids Sequence And Ligation Independent Cloning

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Ligation Independent Cloning (LIC) Workflow Note that times are based on estimates for moving a gene from one plasmid to another. If the source for gene transfer is gDNA add 2 hours to calculation for the traditional cloning method.In this case 15 bp of vectorlogous sequence is used plus a minimum of 18 bp of your template sequence. 5 and 3 primers will have different leader sequences but operate on the same principle (vectorlogous to the first G on 3-5 strand from cut site). For simplicity only the 5 primer is shown here.Ligation Independent Cloning (LIC) is a technique developed in the early 1990s as an alternative to restriction enzymeligase cloning. Inserts are usually PCR amplified and vectors are made linear either by restriction enzyme digestion or by PCR.

Abstract. We describe here a method for sequence- and ligation-independent cloning (SLIC). SLIC uses an exonuclease T4 DNA polymerase to generate single-stranded DNA overhangs in insert and vector sequences.Cited by 159Publish Year 2012Author Mamie Z. Li Mamie Z. Li Stephen J. Elledge Stephen J. ElledgeResults 5-way SLIC SLIC Overview RecA independent recombination is similar to Ligation Independent Cloning (LIC) but without sequence constraints (thus no special vectors needed) via iPCR 10-way SLIC LR reaction (site specific recombination) background from uncleaved vector18.02.2016 If cloning methods had personalities SLIC (sequence- and ligation-independent cloning) would be a true rebel. Not only does this system not use site-specific recombination it also doesnt require a ligation step

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