A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep

Friis, Thor Einar, Stephenson, Sally, Xiao, Yin, Whitehead, Jon and Hutmacher, Dietmar W. (2014) A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep. Tissue Engineering. Part C. Methods, 20 10: 780-789. doi:10.1089/ten.tec.2013.0099


Author Friis, Thor Einar
Stephenson, Sally
Xiao, Yin
Whitehead, Jon
Hutmacher, Dietmar W.
Title A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep
Journal name Tissue Engineering. Part C. Methods   Check publisher's open access policy
ISSN 1937-3384
1937-3392
Publication date 2014-10-01
Year available 2014
Sub-type Article (original research)
DOI 10.1089/ten.tec.2013.0099
Open Access Status Not yet assessed
Volume 20
Issue 10
Start page 780
End page 789
Total pages 10
Place of publication New Rochelle, NY, United States
Publisher Mary Ann Liebert
Language eng
Abstract The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes.
Keyword Full-Length Cdna
Screening Cdna
Messenger-Rna
Animal-Models
Globin Genes
In-Vitro
Bone
Sequence
Pcr
Differentiation
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID LP100200082
C-10-61H
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Mater Research Institute-UQ (MRI-UQ)
 
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Created: Mon, 13 Nov 2017, 01:37:38 EST