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- 2026年01月20日
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文献和实验Inverse PCR for PAC-end sequencing
of the insert. To increase the odds of getting an amplifiable product, we use two different enzymes (used singly!!) per pac end - NlaIII and RsaI for the SP6 end, and NlaIII and HinPI (or HhaI) for the T7 end. Primers PCYPAC2SP6L GCC GTC GAC ATT TAG
Inverse PCR For use with Snyder mTn-lacZ/LEU2 based mutagenesis
1 µl • Use either AciI, AluI, HaeIII, HpaII, RsaI or TaqI for Leu transposon libraries • 37 deg C/ >3 hours (or overnight) • 65 deg C/ 20 min III. Ligations (intramolecular, hopefully)
DOP-PCR Amplification of Whole Genomic DNA and Microchip-Based Capillary Electrophoresis
). In a more demanding protocol, called linker adaptor-PCR, RsaI restricted genomic DNA fragments are ligated to SmaI-cut pUC plasmid. Subsequently, the inserts are amplified by PCR using the universal M13/pUC sequencing and reverse sequencing
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