In the methods paper "A restriction enzyme-PCR-based technique to determine transgene insertion sites by Bryda, E. C. & Bauer, B. A. http://www.ncbi.nlm....pubmed/20013241" they use a Y-linker as an anchor for the second set of primers.
5’ - GTGCAGCCTTGGGTCGCCGTGT/3InvdT/ Y-linker A
3’ – TCACGTCGGAACCCAGCGGCACATGGCAGGAGCGTAAATAGCAAACG Y-linker E
3’ - TGGCAGGAGCGTAAATAGCAAACG Y-linker primer D
3’ - CCAGCGGCACATGGCAGGAGCGTA Y-linker primer G
Since these sequences are listed as the primers required I am a little confused why they would suggest having the primers identical to the Y-linker E sequence and not complementary to it. Am I correct in assuming they expected the reader to get the compliment?
3’ - ACCGTCCTCGCATTTATCGTTTGC
3’ - GGTCGCCGTGTACCGTCCTCGCAT
Seem really strange to make a protocol this complicated, or am I missing something really obvious?