RT @JulianVerdonk: Goodbye Gateway, Gibson, Golden gate, why the hell didn't I know about CPEC?? First cloning attempt, 3 out of 4. Two and…
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Goodbye Gateway, Gibson, Golden gate, why the hell didn't I know about CPEC?? First cloning attempt, 3 out of 4. Two and Three fragment cloning using only proofreading polymerase. Good times in the Bargmann lab @Bas_Bargmann @Asomr1 @kelseyreedplb https:
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@peiferlabunc I had a 12 kb plasmid and no kit worked. What did work was CPEC: make two pairs of primers, one mutagenic, the other in the antibiotic resistance gene. Best if the sizes of the two fragments are 1-2 kb different. Use Phu+ from Thermo. Worked
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PLOS ONE: Circular Polymerase Extension Cloning of Complex Gene Libraries and Pathways http://t.co/JkLnrx56yM
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