Method 5'-Inverse PCR

Method	5'-Inverse PCR
Description	It is necessary to determine the genomic sequence which is adjacent to the inserted trap vector in order to distinguish the homozygote from the heterozygote. inverse PCR is a method to isolate the fragment that is adjacent to the trap vector. First two steps of inverse PCR are common with plasmid rescue. That includes (i) digestion with a suitable restriction enzyme (RE), and (ii) self-ligation. Then, PCR is done with out-sided primers of trap vector.