TABLE 1.
Primer namea | Genomic location (nucleotides) | Nucleotide sequenceb (5′ to 3′) |
---|---|---|
Strand-specific and competitive template primers | ||
T7.5UTR.1 | 1–20 | TAATACGACTCACTATAGGGTTAAAACAGCCTGTGGGTTG |
T7.3UTR.7359 | 7341–7359 | TAATACGACTCACTATAGGGGCACCGTTATCTAGTTCGG |
2C.4365*4774 | 4365–4387* | CATACAGTTCAAGTCCAAATGCC* |
4774–4798 | ATCGAGGTTATCTCCATGTACAGCC | |
3UTR.7359 | 7341–7359 | GCACCGTTATCTAGTTCGG |
REF primers (sense/antisense pairs) | ||
5UTR.1 | 1–20 | TTAAAACAGCCTGTGGGTTG |
5UTR.603 | 584–603 | TCAATTGTCACCATAAGCAG |
5UTR.450 | 450–474 | CCGGCCCCTGAATGCGGCTAATCCT |
1C.2154 | 2134–2154 | TGCGCCTGGTGGTGAGTATGC |
1C.2093 | 2093–2120 | TGTTCTGTGGGTCTGCAATGGCCACAG |
2B.4022 | 3994–4022 | GCCATTGGAATCCCATAATATTGGGACAC |
2A.3307 | 3307–3338 | GGGGCTGTCTACGTAGGCAACTACAGAGTGGT |
2B.4417 | 4393–4417 | CGTGCAAGAGACAGCATACAGGTTC |
2C.4365 | 4365–4387 | CATACAGTTCAAGTCCAAATGCC |
3C.5552 | 5524–5552 | CCATCCTTGTCCACCAACTCTTTAGCGTC |
3C.5479 | 5479–5507 | CCTGGGCCAACCATCCTGATGAATGACCA |
3D.6693 | 6669–6693 | GTTGGTTTCCTTGTGCGAGTATCCG |
3D.6693.82T-C | 6669–6693 | GTTGGTTTCCTCGTGCGAGTATCCGc |
3D.6255 | 6255–6279 | CCCGTATGTCGCATTAGGTATCAAG |
Tag.3DT.C | 7389–poly(A) | CGATCTGAGTCATGTCACTGTTTTTTTTTTTTTTC |
With the exception of Tag.3DT.C, virus-specific primer names include the viral gene designation and the number of the first nucleotide on the 5′ end of the primer. In primer 2C.4365*4774, the location of a deletion that was engineered into the primer is indicated (*).
Primers 5UTR, 5UTR.603, 5UTR.450, 1C.2154, 2B.4022, and 2B.4417 were based on the published sequence of pNI4. All remaining primer sequences were based on sequence information obtained from CVB1T (unpublished data). The Tag sequence in primer Tag.3DT.C is a nonviral sequence appended to the 5′ end to facilitate amplification of the 3′ end of the virus and is italicized. T7 promoter sequences in T7.5UTR.1 and T7.3UTR.7359 are underlined.
This primer has the same sequence as 3D.6669-93 but with a T-to-C transition at nucleotide 6682 (shown in boldtype).