›› 2011, Vol. 38 ›› Issue (3): 80-83.

• 生物技术 • Previous Articles     Next Articles

Sperm RNA Extraction from Dairy Cattle

WANG Hui-e1，2，SUN Xian-ying2，GAO Qing-hua1，2，HAN Chun-mei2，SHEN Bo2，WU Yan-feng2   

  1. 1.College of Animal Science and Technology，Tarim University，Alar 843300,China; 2.Key Laboratory of Tarim Animal Husbandry Science and Technology，Xinjiang Production and Construction Groups，Tarim University，Alar 843300,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-03-20 Published:2011-03-20
  • Contact: GAO Qing-hua

Abstract: Isolation of high-quality RNA is important for the study of sperm gene expression. Frozen straw semen from three bulls were purified by swimming up. The laminar analysis column method and Trizol single-step method were combined for RNA extraction from swimming-up bovine frozen-thaw sperm，and non-swimming up sperm as control. Sperm total RNA was analyzed with UV spectrophotometer，RT-PCR and garose gel electrophoresis. Results showed that the bovine sperm swimming-up from frozen straws were not contaminated by somatic cells，and high-quality total RNA was obtained in about 2 h. The value of D260/280 was 1.8，and the ≥200 bp total RNA extracted from bovine sperm yielded 0.92 μg/107sperm.Following RT-PCR，clear bands of SRY，LEP，and RPS23 of sperm cDNA were shown in a garose gel electrophoresis respectively，not a band of 18S rRNA. The purity and integrality of total RNA isolated from the bovine sperm of frozen straw with the method of combined the laminar analysis column with Trizol single-step were significantly satisfactory for the demands of molecular biological experiments.

Key words: bovine; sperm; total RNA; RT-PCR

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