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The enhancement of the sensitivity of a pcr-based avian sex determination assay.

Reproduction, Fertility and Development.. 2007-12;  20(1):214-215
C. Valentin, R. West, C. Herr.
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摘要

For over a decade it has been possible to externally sex monomorphic birds using PCR. A major drawback of the protocol developed by Richard Griffiths is that DNA from at least 20 000 cells is needed (Griffiths et al. 1996 Proc. Royal Soc. London B 263, 1249–1254). Our study attempted to decrease the number of cells required. A sequence within the chromobox-helicase-DNA-binding (CHD) gene, located on the sex chromosomes of all avian species, was amplified. The sequence lengths were 362 and 354 base pairs for the CHD-W and CHD-Z, respectively. The polyacrylamide gel electrophoresis (PAGE) purified primers used were 5′-TCTGCATCGCTAAATCCTTT-3′ and 5′-CTCCCAAGGATGAGRAAYTG-3′ (2.5 μm) (IDT, Inc., San Jose, CA,... More

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