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Spermatozoa input concentrations and RNA isolation methods on RNA yield and quality in bull (Bos taurus).

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Title Spermatozoa input concentrations and RNA isolation methods on RNA yield and quality in bull (Bos taurus).
Not Available
 
Creator Parthipan S
Selvaraju S
Somashekar L
Kolte AP
Arangasamy A
Ravindra JP
 
Subject Bull
Spermatozoa
RNA isolation
RNA yield
RNA quality
 
Description Not Available
Sperm RNA can be used to understand the past spermatogenic process, future successful fertilization, and embryo development. To study the sperm RNA composition and function, isolation of good quality RNA with sufficient quantity is essential. The objective of this study was to assess the influence of sperm input concentrations and RNA isolation methods on RNA yield and quality in bull sperm. The fresh semen samples from bulls (n = 6) were snap-frozen in liquid nitrogen and stored at −80 °C. The sperm RNA was isolated using membrane-based methods combined with TRIzol (RNeasy + TRIzol and PureLink + TRIzol) and conventional methods (TRIzol, Double TRIzol, and RNAzol RT). Based on fluorometric quantification, combined methods resulted in significantly (P < 0.05) higher total RNA yields (800–900 ng/30–40 × 106) as compared with other methods and yielded 20 to 30 fg of RNA/spermatozoon. The quality of RNA isolated by membrane-based methods was superior to that isolated by conventional methods. The sperm RNA was observed to be intact as well as fragmented (50–2000 bp). The study revealed that the membrane-based methods with a cocktail of lysis solution and an optimal input concentration of 30 to 40 million sperm were optimal for maximum recovery of RNA from bull spermatozoa.
Not Available
 
Date 2018-11-23T07:13:59Z
2018-11-23T07:13:59Z
2015-08-01
 
Type Research Paper
 
Identifier Parthipan S, Selvaraju S, Somashekar L, Kolte AP, Arangasamy A and Ravindra JP. 2015. Spermatozoa input concentrations and RNA isolation methods on RNA yield and quality in bull (Bos taurus). Analytical Biochemistry, 482:32-39.
0003-2697
http://krishi.icar.gov.in/jspui/handle/123456789/12942
 
Language English
 
Relation Not Available;
 
Publisher Elsevier