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Novel extraction method of genomic DNA suitable for long-fragment amplification from small amounts of milk.

Authors :
Liu, Y. F.
Gao, J. L.
Yang, Y. F.
Ku, T.
Zan, L. S.
Source :
Journal of Dairy Science. Nov2014, Vol. 97 Issue 11, p6804-6809. 6p.
Publication Year :
2014

Abstract

Isolation of genomic DNA is a prerequisite for assessment of milk quality. As a source of genomic DNA, milk Somatic cells from milking ruminants are practical, animal friendly, and cost-effective sources. Extracting DNA from milk can avoid the stress response caused by blood and tissue sampling of cows. In this study, we optimized a novel DNA extraction method for amplifying long (>1,000 bp) DNA fragments and used it to evaluate the isolation of DNA from small amounts of milk. The techniques used for the separation of milk omatic cell were explored and combined with a sodium dodecyl sulfate (SDS)-phenol method for optimizing DNA extraction from milk. Spectrophotometry was used to determine the concentration and purity of the extracted DNA. Gel electrophoresis and DNA amplification technologies were used for to determine DNA size and quality. The DNA of 112 cows was obtained from milk (samples of 13 ± 1 mL) and the corresponding optical density ratios at 260:280 nm were between 1.65 and 1.75. Concentrations were between 12 and 45 µg/µL and DNA size and quality were acceptable. The pecific PCR amplification of 1,019- and 729-bp bovine DNA fragments was successfully carried out. This novel method can be used as a practical, fast, and economical mean for long genomic DNA extraction from a small amount of milk. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00220302
Volume :
97
Issue :
11
Database :
Academic Search Index
Journal :
Journal of Dairy Science
Publication Type :
Academic Journal
Accession number :
99080888
Full Text :
https://doi.org/10.3168/jds.2014-8066