| Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia. | |
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MedLine Citation:
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PMID: 23199708 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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OBJECTIVE: To construct rapidly a full-length cDNA library from nanogram amounts total RNA of Giardia lamblia (G. lamblia) trophozoites stocked in RNA stabilization reagent. METHODS: Total RNA of Giardia was extracted using Trizol reagent. A full-length cDNA library of G. lamblia trophozoites was constructed by a long-distance PCR (LD-PCR) method. The recombinant rate and the coverage rate of full-length clones of the library were evaluated. The inserted fragments were identified and sequenced by PCR amplification. RESULTS: The titer of cDNA library was 3.85 × 10(7) pfu/mL. The length of inserted fragments ranged from 0.4 to 2.5 kb, and the recombination efficiency accounted for 100% (20/20). The coverage rate of full-length clones is high (17/20). CONCLUSIONS: The RNA stabilization reagent may be used to fix the cells and prevent the RNA in cells even though delivered under normal atmospheric temperature. The long-distance PCR can be used to construct a full-length cDNA library rapidly and it needs less RNA than the traditional method from mRNA. |
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Authors:
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Jun-Li Guo; Jie Jiang; Wen-Yu Zheng; Ming-Luan Li; Xi-Feng Tian; Xian-Min Feng; Yue-Hua Wang; Xiao-Hong Ju; Yue-Qiong Kong |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Asian Pacific journal of tropical medicine Volume: 5 ISSN: 1995-7645 ISO Abbreviation: Asian Pac J Trop Med Publication Date: 2012 Dec |
Date Detail:
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Created Date: 2012-12-03 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101533720 Medline TA: Asian Pac J Trop Med Country: Singapore |
Other Details:
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Languages: eng Pagination: 931-4 Citation Subset: IM |
Copyright Information:
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Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved. |
Affiliation:
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School of Laboratory Medicine, Jilin Medical College, Jilin 132013, PR China; Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical College, Haikou 571199, PR China. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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