Table 1 PCR Primers used in this study
Name | Sequences (5' - 3') | Comments |
|---|---|---|
P450F1 | TGAAAATTAACCAGCCACCAT | Gene and cDNA cloning |
P450R1 | TTATATCATTTGTGAAAGC GACAAG | Gene and cDNA cloning |
P450F2 | TGCATGGACTGATGCAGAC | Expression detection in gel |
P450R2 | CTCCTCATTCATGTCCAACTC | Expression detection in gel |
P450F3 | TGGAGTTGCTCAGCAGCCATA | Internal primer for DNA sequencing |
P450R3 | TATATGGAA GCCATCGACTGTG | Internal primer for DNA sequencing |
5'UTRF1 | TGCAGCTTCATATCTTGGGTTTTCTC | 5'UTR cloning (1000 bp) |
5'UTRR3 | GGTGGCTGGTT AATTTTCAGTATTCAG | 5'UTR cloning (1000 bp) |
3'UTRF1 | ATATAATCTCTGTATCATTGCCAACTGAG | 3'UTR cloning (600 bp) |
3'UTRR1 | ATGCTAAACATCAAATCGAATACTCTCA | 3'UTR cloning (600 bp) |
SYBR-P450F1 | TGCATGGACTGATGCAGAC | Real-time qPCR for gene expression |
SYBR-P450R1 | CTCCTCATTCATGTCCAACTC | Real-time qPCR for gene expression |
SYBR-P450F2 | CCAACATCAAAGCTATATCTTTG | Real-time qPCR for pre-mRNA splicing |
SYBR-P450R2 | TGGAGGCTGCCATCAT ATC | Real-time qPCR for pre-mRNA splicing |
P450-5'RACE GSP2 | CGATACTTTTTGGATAGAGCTTGTAGAGC | First 5'RACE |
P450-5'RACE GSP3 | CGACAAGCTCCCTAACATATGCATATT | Second 5'RACE |
P450-3'RACE GSP1 | CGAGCAACTTGTTCATTGCTTTGAT | First 3'RACE |
P450-3'RACE GSP2 | CGAAGA TAACATGTTGGCAAGTGAGTT | Second 3'RACE |
SYBR-P450B 5'UTRF1 | ACTGACTTCTAGTTTAAATTTTTCTT | Real-time qPCR for 5'RACE |
SYBR-P450B 5'UTRR1 | TGGCTGGTTAATTTTCAGTATT | Real-time qPCR for 5'RACE |
SYBR-P450B 3'UTRF1 | ATATAATCTCTGTATC ATTGCCAA | Real-time qPCR for 3'RACE |
SYBR-P450B 3'UTRR1 | ACTACCAGATGAAAATCATTAAAT | Real-time qPCR for 3'RACE |
P450-5'RACE GSP3 | CGACAAGCTCCCTAACATATGCATATT | 5' primer extension |
P450-3'RACE GSP2 | CGAAGA TAACATGTTGGCAAGTGAGTT | 3' primer extension |
-300UF | AGTCATATATGACTTAGCAAGAGAACTCCAC | PCR for pre-mRNA splicing at -300 TSS |
-140UF | AACCGCACCTTATC CTCTTCACAA | PCR for pre-mRNA splicing at -140 TSS |
-60UF | TGAAACCACCCAAGAACTTCAAAAC | PCR for pre-mRNA splicing at -60 TSS |
++70UR | CATAAAGCATAAAGCGTTATTATTCATATTT | PCR for pre-mRNA splicing at ++70 poly(A) site |
++200UR | CTAAGTTCCATATTCT TCCTTCTTAAGTTCA | PCR for pre-mRNA splicing at ++200 poly(A) site |
++260UR | ACTCAGAACTAAGTTTTATTTCACTTGATCAA | PCR for pre-mRNA splicing at ++260 poly(A) site |