-transparent.png){kind=logo}
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.
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
April 2010,
BMC molecular biology,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
August 2003,
Analytical biochemistry,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
October 2000,
Cancer,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
September 2001,
Cancer letters,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
May 1990,
Journal of the National Cancer Institute,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
January 1993,
Methods in enzymology,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
December 1996,
Analytical biochemistry,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
July 1993,
Analytical biochemistry,
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
January 1998,
Methods in molecular biology (Clifton, N.J.),
I Kagami, and
H Mizunuma, and
S Miyamoto, and
Y Ibuki, and
T Uchida
January 2001,
Oncology,
