Table 1.
Primers for specific amplification of Δ133p53 isoforms mRNAs by nested RT-PCR. The specific region (exon or intron) that each of the primers target is indicated (Ex: exon; Int: intron), the sequences corresponding to the exon junction are underlined. (F): Forward, (R): Reverse. PCR fragment sizes (bp) corresponding to p53 isoforms are also highlighted. Quality of reverse-transcription is assessed by quantitative RT-PCR amplification (SybrGreen) of actin and p53 mRNAs (primers are included)
| p53 mRNA variant |
PCR | Primer name and targeted region | 5’ – 3’ sequence |
|---|---|---|---|
| All p53 mRNA | For6 (F) (Ex6) | TTGCGTGTGGAGTATTTGGAT | |
| Rev7 (R) (Ex7) | TGTAGTGGATGGTGGTACAGTCAGA | ||
| Δ133p53α | 1st | D133F1 (F) (Int4) | TAGACGCCAACTCTCTCTAG |
| Rev10 (R) (Ex10) | CTT CCC AGC CTG GGC ATC CTT G | ||
| 2nd
(670bp) |
D133F2 (F) (Int4) | ACT CTG TCT CCT TCC TCT TCC TAC AG | |
| RDNp53 (R) (Ex9/Ex10) | CTC ACG CCC ACG GAT CTG A | ||
| Δ133p53β | 1st | D133F1 (F) (Int4) | TAGACGCCAACTCTCTCTAG |
| Rev10 (R) (Ex10) | CTT CCC AGC CTG GGC ATC CTT G | ||
| 2nd
(700bp) |
D133F2 (F) (Int4) | ACT CTG TCT CCT TCC TCT TCC TAC AG | |
| p53β (R) (Ex9β) | TCA TAG AAC CAT TTT CAT GCT CTC TT | ||
| Δ133p53γ | 1st | D133F1 (F) (Int4) | TAGACGCCAACTCTCTCTAG |
| Rev10 (R) (Ex10) | CTT CCC AGC CTG GGC ATC CTT G | ||
| 2nd
(670bp) |
D133F2 (F) (Int4) | ACT CTG TCT CCT TCC TCT TCC TAC AG | |
| p53γ (R) (Ex9/Ex9γ) | TCGTAAGTCAAGTAGCATCTGAAGG | ||
| actin | actin (F) | ATCTGGCACCACACCTTCTACAATGAGCTGCG | |
| actin (R) | CGTCATACTCCTGCTTGCTGATCCACATCTGC | ||