Appendix I: Oligo Sequences
Appendix I: Oligo Sequences
| Primer Name | Primer Sequence | Expected PCR Product Length (bp) |
| T7 Fragment 1 Forward | TATAACGTTTTTGAACACACATGAACAAGG
TCTCACAGTGTACGGACCTAAAGTTCCCCC |
9999 |
| T7 Fragment 1 Reverse | ATTACGCGATGACAGTAGACAACCTTTCCG | |
| T7 Fragment 2 Forward | TGCAGCAATACCGGAAAGGTTGTCTACTGT | 10,000 |
| T7 Fragment 2 Reverse | ATATGTCTCCTCATAGATGTGCCTATGTGG | |
| T7 Fragment 3 Forward | ACTTGTGACTCCACATAGGCACATCTATGA | 10,000 |
| T7 Fragment 3 Reverse | GAATAACCTGAGGGTCAATACCCTGCTTGT | |
| T7 Fragment 4 Forward (F4F)* | GACATGATGGACAAGCAGGGTATTGACCCT | |
| T7 Fragment 5 Reverse (F5R)* | ACATTCAAATATGTATCCGCTCATGAGACAA
GGGACACAGAGAGACACTCAAGGTAACAC |
|
| T7 D540H Forward | CACagtcgttcagtacctaatgcaatcatgg | 846, with F5R |
| T7 D540H Reverse | cattaggtactgaacgactGTGtgcaatattc | 9213, with F4F |
| T7 D540S Forward | TCCagtcgttcagtacctaatgcaatcatgg | 846, with F5R |
| T7 D540S Reverse | cattaggtactgaacgactGGAtgcaatattc | 9213, with F4F |
| T7 S541D Forward | gacGATcgttcagtacctaatgcaatcatgg | 843, with F5R |
| T7 S541D Reverse | cattaggtactgaacgATCtgatgcaatattc | 9216, with F4F |
| T7 S541K Forward | gacAAAcgttcagtacctaatgcaatcatgg | 843, with F5R |
| T7 S541K Reverse | cattaggtactgaacgTTTtgatgcaatattc | 9216, with F4F |
| V544R Forward | AGAcctaatgcaatcatggtggagaacgag | 837, with F5R |
| V544R Reverse |
cattaggTCTtgaacgactgtctgcaatattc |
9222, with F4F |
| V544H Forward | CACcctaatgcaatcatggtggagaacgag | 837, with F5R |
| V544H Reverse | cattaggGTGtgaacgactgtctgcaatattc | 9222, with F4F |
| pRS315 P1 Forward | TGTCTCATGAGCGGATACATATTTGAATGT | 713 |
| pRS315 P1 Reverse | CCTTGTTCATGTGTGTTCAAAAACGTTATA | |
| pRS315 P2 Forward | GCATGTGTCAGAGGTTTTCACCG | 564 |
| pRS315 P2 Reverse | CGCGTTTCGGTGATGACGG | |
| pRS315 P3 Forward | GGGTAGACGAAACTATATACGC | 1755 |
| pRS315 P3 Reverse | GCGTATATAGTTTCGTCTACCC | |
| F1-F2 Forward | GCGAATCAAGGTGACTTCCTC | 900 |
| F1-F2 Reverse | GGAGTTCCTATGGATAGGTGTTCAGG | |
| F2-F3 Forward | GAGGTTTCCACTGAGGACGAAG | 85 |
| F2-F3 Reverse | CCTTTGGTAACCATGTCCTCGG | |
| F3-F4 Forward | CTCCGTGAGTCCATGCAGTT | 210 |
| F3-F4 Reverse | GGCGATCCCCCTAGAGTCTT | |
| F4-F5 Forward | GCCCCTGTGTGTTCTCGTTA | ~500 |
| F4-F5 Reverse | GTGTGGTGACTCACTGTCGT |
*Sequence of T7 Fragment 4 Reverse and T7 Fragment 5 Forward varies based on the mutation