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Multiplex standardized RT-PCR for expression analysis of many genes in small samples.

Abstract：

:Standardized RT-PCR (StaRT-PCR) enables numerical quantification as well as intra- and inter-laboratory comparison of gene expression. Multiplex StaRT-PCR, using two rounds of amplification, was conducted on Stratagene Universal Reference RNA. In the first round, cDNA, competitive template (CT) mix, and primers for up to 96 genes were amplified for varying numbers of cycles. Next, products from round one were diluted, combined with primers for one gene, and amplified for an additional 35 cycles. No additional cDNA or CT mix was added. Expression values obtained by uniplex and multiplex StaRT-PCRs were highly correlated (R=0.993, p<0.001). Products from round one could be diluted as much as 100,000-fold and still be quantified following round two amplification. Thus, using multiplex StaRT-PCR, 96 genes were measured in the same amount of cDNA typically used to measure one gene with uniplex StaRT-PCR. Multiplex StaRT-PCR was also used to measure 18 genes in the fine needle biopsy of a primary lung carcinoma.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Crawford EL,Warner KA,Khuder SA,Zahorchak RJ,Willey JC

doi

10.1016/S0006-291X(02)00243-7

subject

Has Abstract

pub_date

2002-04-26 00:00:00

pages

509-16

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(02)00243-7

journal_volume

293

pub_type

杂志文章

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