Table 1 Primers for PST-PCR. Abbreviations: Tm, melting temperature; CG% (percentage of C and G) calculated for the core sequence (3′-end palindrome and 10 degenerate positions) at primer concentration of 0.5 µM in the presence of 2 mM Mg2+; for the tail primer, calculations were made for 0.25 µM primer in absence of Mg2+; LC (%) - Linguistic Complexity calculated for 6-nt restriction sites.
ID | Sequence (5′-3′) | Restriction site | Tm (°C) | CG (%) | LC (%) |
|---|---|---|---|---|---|
Tail primer (for second round PCR) | |||||
5600 | GTTGCGGCAGGTCCTCACC | — | 69.1 | 68.1 | 89 |
PST primers (for first round PCR) | |||||
5601 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGACGTC | AatII | 49.3 | 56.3 | 100 |
5602 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnAACGTT | AclI | 46.1 | 43.8 | 100 |
5603 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnTTCGAA | AsuII | 45.6 | 43.8 | 100 |
5604 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnTGGCCA | BalI | 51.0 | 56.3 | 100 |
5605 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGGATCC | BamHI | 48.3 | 56.3 | 100 |
5606 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnTGATCA | BclI | 44.7 | 43.8 | 100 |
5326 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnAGATCT | BglII | 43.7 | 43.8 | 100 |
5607 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnATCGAT | ClaI | 44.8 | 43.8 | 89 |
5608 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGAATTC | EcoRI | 43.7 | 43.8 | 100 |
5609 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGATATC | EcoRV | 42.3 | 43.8 | 89 |
5610 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnAAGCTT | HindIII | 45.5 | 43.8 | 100 |
5611 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGTTAAC | HpaI | 43.6 | 43.8 | 100 |
5612 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGGTACC | KpnI | 48.2 | 56.3 | 100 |
5613 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnCCATGG | NcoI | 49.1 | 56.3 | 100 |
5614 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGCTAGC | NheI | 49.3 | 56.3 | 89 |
5615 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnCACGTG | PmaCI | 50.2 | 56.3 | 100 |
5616 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnCTGCAG | PstI | 49.7 | 56.3 | 100 |
5617 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnCAGCTG | PvuII | 49.7 | 56.3 | 100 |
5618 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGAGCTC | SacI | 48.8 | 56.3 | 100 |
5619 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGTCGAC | SalI | 49.3 | 56.3 | 100 |
5620 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnAGTACT | ScaI | 43.7 | 43.8 | 100 |
5621 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnGCATGC | SphI | 50.9 | 56.3 | 89 |
5622 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnAGGCCT | StuI | 50.1 | 56.3 | 100 |
5327 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnTCTAGA | XbaI | 43.1 | 43.8 | 100 |
5623 | GTTGCGGCAGGTCCTCACCnnnnnnnnnnCTCGAG | XhoI | 48.6 | 56.3 | 100 |
