RRC ID 38852
著者 Kagami I, Mizunuma H, Miyamoto S, Ibuki Y, Uchida T.
タイトル Quantification of estrogen receptor messenger RNA by quantitative polymerase chain reaction using internal standard fragment.
ジャーナル Biochem Biophys Res Commun
Abstract A simple and reliable polymerase chain reaction-based method for quantifying human and rat estrogen receptor (ER)-mRNA is described. This method involves co-amplification of cDNA transcribed from sample RNA with an internal standard to reduce tube to tube amplification variations. Human and rat internal standards were synthesized by insertion of a DNA fragment near the midportion of human and rat target ER cDNA molecules. After co-amplification, both products of target ER cDNA and internal standard were separated on agarose gel by electrophoresis and transferred onto a membrane filter. The radioactivity hybridized with 32P-labeled ER cDNA was counted. The ER mRNA content was calculated by linear regression analysis after obtaining a logarithm of ratios between the sample and the internal standard radioactivity. The coefficient of variation of this assay was 18.8%. An absolute amount of ER mRNA in less than 10(4) cultured cells could be measured.
巻・号 228(2)
ページ 358-64
公開日 1996-11-12
DOI 10.1006/bbrc.1996.1666
PII S0006-291X(96)91666-6
PMID 8920919
MeSH Animals Base Sequence DNA Primers DNA, Complementary Endometrial Neoplasms / metabolism Endometrium / metabolism Estradiol / pharmacology Female Humans Male Mutagenesis, Insertional Orchiectomy Polymerase Chain Reaction / methods* RNA, Messenger / analysis* Rats Rats, Wistar Receptors, Estrogen / biosynthesis* Recombinant Proteins / biosynthesis Reference Standards Reproducibility of Results Transcription, Genetic / drug effects Uterus / drug effects Uterus / metabolism*
IF 2.985
引用数 5
WOS 分野 BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 SKN(RCB0513) HHUA(RCB0658) HOS(RCB0992) ROS-17/2.8-5(RCB0462)