ogous to PCR products amplified by Taq DNA polymerase. TAcloning is therefore recommended. For blunt-end cloning, the fragments need to be polished by Klenow enzyme or T4 DNA polymerase.

11.8 Sequencing

Sequencing with the TripleMaster Polymerase Mix is not recommended due to the inherent 35 exonuclease activity.

11.9 Primer sequences for long targets

universal forward primer (in the control primer mix) :
5-CTGATGAGTTCGTGTCCGTACAACTGGCGTAATC-3 reverse primers: 40 kb: 5-TAATGCAAACTACGCGCCCTCGTATCACATGG-3 (in the control primer mix) 35 kb: 5-ATTATGTCGGTGATACTTCGTCGCTGTCTC-3 30 kb: 5-GAAAGTTATCGCTAGTCAGTGGCCTGAAGAGACG-3 20 kb: 5-GTGCACCATGCAACATGAATAACAGTGGGTTATC-3 10 kb: 5-ATACGCTGTATTCAGCAACACCGTCAGGAACACG-3 universal forward primer for human tPA gene: 5-CCTTCACTGTCTGCCTAACTCCTTCGTGTGTTCC-3 reverse primer for human tPA gene: 15 kb: 5- ACTGTGCTTCCTGACCCATGGCAGAAGCGCCTTC-3 18 kb: 5-GCAGGGGTGCTGCAGAACTCTGAGCTGTACTTCC-3 22 kb: 5-GATGCGAAACTGAGGCTGGCTGTACTGTCTC-3 24 kb: 5-TGTCTCCAGCACACAGCATGTTGTCGGTGAC-3
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