医学英语分子生物学实验,英译汉Compare the cDNA sequence of 4 variants,there
医学英语分子生物学实验,英译汉
Compare the cDNA sequence of 4 variants,there are only differences between exon3 and exon7,thus design specific primer for these two exons could differentiate 4 variants.
1.total RNA extraction
2.RT-PCR:
primer for exon3:Tm:about 52°C
sense primer:5' GATTGGACAGGCTGGACT 3'
anti-sense primer:5' AGAAGTGGTGGCAGTGATT 3'
predicted results:288bp product in Nurr1-a,b,c; no amplified product in Nurr1-d.
primer for exon7:Tm:58-59°C
sense primer:5' CCAAAGCCGACCAAGACC 3'
anti-sense primer:5' ATTATTGCTGGCGGTGGC 3'
predicted results:472bp product in Nurr1-a,d; 418bp product in Nurr1-b; 351bp product in Nurr1-c.
3.result analysis:
Nurr1-a Nurr1-b Nurr1-c Nurr1-d
amplication for exon 3 288bp 288bp 288bp no product
amplication for exon 7 472bp 418bp 351bp 472bp
4.
Problem:RefSeq for Nurr1-b,c,d have been permanently suppressed because of insufficient support.