奶牛圆形、长形、成熟精子细胞微量RNA制备和含量分析

doi:doi: 10.11843/j.issn.0366-6964.2016.08.004

Abstract

Abstract:

To explore the method to capture dairy cattles round and elongated spermatids and build the stable system extracting sperm RNA and analyzing the RNA quantity of spermatogenic cells，the round and elongated spermatids were captured by laser capture microdissection (LCM) combined with the technology of frozen section，and sperm samples were obtained using the method of sperm floating from epididymids.Compared the reported sperm RNA preparation methods，a better RNA preparation result was obtained from dairy cattle sperm using RNeasy Kit，and the better RNA preparation results were also obtained from round and elongated spermatids using PicoPure RNA Isolation Kit.Besides，the removal results of contaminated somatic cells and genomic DNA were tested by reverse transcription polymerase chain reaction（RT-PCR）.The one-way analysis of variance showed that the round and elongated spermatids were successfully captured by LCM，the high quality RNA was extracted from sperm using RNeasy Kit，and the RNA quantity of each sperm was 0.023-0.025 pg.The RNA quantity decreased by 750-fold from round to elongated spermatids.The exploration that capture spermatogenic cells from complex tissues and extract RNA from less RNA containing cells will provide an important reference for the study on mechanism of spermatogenesis and spermiogenesis.

CLC Number:

S814

REN Xiao-xia，CHEN Xiao-li，LI Xiu-ling，WANG Yan，QIN Tong，WANG Dong. The Preparation and Quantity Analysis of Trace RNA from Round Spermatids，Elongated Spermatids and Mature Sperm of Dairy Cattle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(8): 1546-1554.

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The Preparation and Quantity Analysis of Trace RNA from Round Spermatids，Elongated Spermatids and Mature Sperm of Dairy Cattle

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