cDNA libraries from single human preimplantation embryos

Genomics
J AdjayeM Monk

Abstract

In this paper, the construction, evaluation, and application of cDNA libraries from eight unfertilized oocytes and single four-cell-, seven-cell-, and blastocyst-stage embryos are described. Rapid, reproducible, and efficient procedures for the construction of PCR-based cDNA libraries from fewer than 10 cells were first developed in small populations of fibroblast cells. The human embryo libraries display complexities sufficient (between 10(5) and 10(6) clones) to represent the entire active gene population at these early stages of human development. The ubiquitous cytoskeletal elements, beta-actin, keratin-18, and alpha-tubulin, were detected at the expected frequency. Sequencing of consecutively picked random clones, without selection, showed the presence of a variety of sequences, such as the human transposable element, LINE-1 and Alu repeat sequences, housekeeping genes, and tissue-specific genes, such as alpha-globin and FMR-1. In addition to cDNAs corresponding to known ESTs (expressed sequence tags) in the GenBank and dbEST databases, a high proportion of novel sequences were detected. Applications of the libraries to several areas of interest, such as expression of CpG-island-containing "tissue-specific" genes, developm...Continue Reading

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Citations

Apr 12, 2001·International Journal of Experimental Pathology·R J ByersA J Freemont
Apr 15, 2011·Genetics and Molecular Research : GMR·Y P MaG W Ren
Jan 7, 2004·Development, Growth & Differentiation·Wen Yong LiQing Xuan Chen
Mar 27, 2003·Molecular Reproduction and Development·Anthony D MetcalfeDaniel R Brison
Apr 14, 2009·Molecular Reproduction and Development·Alexei V Evsikov, Caralina Marín de Evsikova
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Apr 23, 2005·Human Reproduction·Maria D SeraficaAlan O Trounson
Feb 15, 2002·Human Fertility : Journal of the British Fertility Society·Joanna HurleyJames Adjaye

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