Literature DB >> 22992930

Isolating mRNA and small noncoding RNAs from human sperm.

Robert J Goodrich1, Ester Anton, Stephen A Krawetz.   

Abstract

The isolation of spermatozoal RNA is a challenging procedure due to the intrinsic heterogeneous population of cells present in the ejaculate and the small quantity of RNA present in sperm. The transcriptome of these gametes includes a wide variety of messenger RNAs (mRNAs), small noncoding RNAs (sncRNAs), and highly fragmented ribosomal RNAs (rRNAs).The protocol described in this chapter to isolate both the mRNA and sncRNA fractions represents years of development towards automation. It combines a guanidinium thiocyanate-phenol-chloroform-based methodology to reduce the content of DNA and a column-based system. Both manual and semi-automated options are described, with preference given to automation for consistent results. A novel quality control procedure has been developed to assess the integrity and purity of the entire population of isolated mRNAs due to the absence of intact rRNAs.

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Year:  2013        PMID： 22992930     DOI： 10.1007/978-1-62703-038-0_33

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN： 1064-3745


  25 in total

Review 1.  The presence, role and clinical use of spermatozoal RNAs.

Authors:  Meritxell Jodar; Sellappan Selvaraju; Edward Sendler; Michael P Diamond; Stephen A Krawetz
Journal:  Hum Reprod Update       Date:  2013-07-14       Impact factor: 15.610

2.  Rapid method for the isolation of mammalian sperm DNA.

Authors:  Haotian Wu; Matthew K de Gannes; Gianna Luchetti; J Richard Pilsner
Journal:  Biotechniques       Date:  2015-06-01       Impact factor: 1.993

3.  Zika virus persistence in the male macaque reproductive tract.

Authors:  Erin E Ball; Patricia A Pesavento; Koen K A Van Rompay; M Kevin Keel; Anil Singapuri; Jose P Gomez-Vazquez; Dawn M Dudley; David H O'Connor; Meghan E Breitbach; Nicholas J Maness; Blake Schouest; Antonito Panganiban; Lark L Coffey
Journal:  PLoS Negl Trop Dis       Date:  2022-07-05

4.  High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing.

Authors:  Andrew P Georgiadis; Archana Kishore; Michelle Zorrilla; Thomas M Jaffe; Joseph S Sanfilippo; Etta Volk; Aleksandar Rajkovic; Alexander N Yatsenko
Journal:  J Urol       Date:  2014-08-01       Impact factor: 7.450

5.  Evaluation of the effectiveness of semen storage and sperm purification methods for spermatozoa transcript profiling.

Authors:  Shihong Mao; Robert J Goodrich; Russ Hauser; Steven M Schrader; Zhen Chen; Stephen A Krawetz
Journal:  Syst Biol Reprod Med       Date:  2013-07-19       Impact factor: 3.061

6.  What human sperm RNA-Seq tells us about the microbiome.

Authors:  Grace M Swanson; Sergey Moskovtsev; Clifford Librach; J Richard Pilsner; Robert Goodrich; Stephen A Krawetz
Journal:  J Assist Reprod Genet       Date:  2020-01-04       Impact factor: 3.412

7.  An Efficient Approach for RNA Extraction from Boar Sperm and Seminal Plasma.

Authors:  Vanmathy R Kasimanickam; Ramanathan K Kasimanickam
Journal:  Bio Protoc       Date:  2019-07-05

8.  Testicular "Inherited Metabolic Memory" of Ancestral High-Fat Diet Is Associated with Sperm sncRNA Content.

Authors:  Luís Crisóstomo; Matthieu Bourgery; Luís Rato; João F Raposo; Rachel L Batterham; Noora Kotaja; Marco G Alves
Journal:  Biomedicines       Date:  2022-04-15

9.  Stability, delivery and functions of human sperm RNAs at fertilization.

Authors:  Edward Sendler; Graham D Johnson; Shihong Mao; Robert J Goodrich; Michael P Diamond; Russ Hauser; Stephen A Krawetz
Journal:  Nucleic Acids Res       Date:  2013-03-06       Impact factor: 16.971

10.  RNA profiles of porcine embryos during genome activation reveal complex metabolic switch sensitive to in vitro conditions.

Authors:  Olga Østrup; Gayla Olbricht; Esben Østrup; Poul Hyttel; Philippe Collas; Ryan Cabot
Journal:  PLoS One       Date:  2013-04-29       Impact factor: 3.240
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