| An alternative approach to synthesize cDNA bypassing traditional reverse transcription. | |
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MedLine Citation:
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PMID: 18228164 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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cDNAs of certain target genes are difficult to obtain by traditional reverse transcription. Herein we describe a novel method to synthesize cDNA based upon the use of the class IIS restriction enzymes. Briefly, the exons of a certain gene are separately PCR-amplified, each using the primers containing a recognition sequence of a certain class IIS restriction enzyme. All the fragments are restricted using the enzyme(s), resulting in the cohesive end of each exon that is complementary to the one in its adjacent exon. Then the fragments can be assembled together in their naturally occurring order. We successfully applied this method to acquire the coding sequence of Hoxa7 gene. This approach is simple, highly efficient, less error prone and cost-effective, and can also be used to fuse different PCR-fragments from distinct genes to create a chimeric gene or to perform site-directed mutagenesis. |
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Authors:
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Xiao-Xia Li; Fang Zheng; Yan-Li Jiao; Gang Guo; Bao-Li Wang; Zhi Yao |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-01-29 |
Journal Detail:
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Title: Molecular biotechnology Volume: 39 ISSN: 1073-6085 ISO Abbreviation: Mol. Biotechnol. Publication Date: 2008 Jul |
Date Detail:
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Created Date: 2008-06-10 Completed Date: 2008-08-21 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9423533 Medline TA: Mol Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 201-6 Citation Subset: IM |
Affiliation:
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Key Research Lab of Hormone and Development Affiliated to the Ministry of Health, Tianjin Medical University, Tianjin 300070, PR China. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Base Sequence Cloning, Molecular Cost-Benefit Analysis DNA Primers DNA, Complementary / genetics* Polymerase Chain Reaction Transcription, Genetic* |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/DNA, Complementary |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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