Document Detail

Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia.
MedLine Citation:
PMID:  23199708     Owner:  NLM     Status:  In-Data-Review    
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.
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:
Type:  Journal Article    
Journal Detail:
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:
Created Date:  2012-12-03     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101533720     Medline TA:  Asian Pac J Trop Med     Country:  Singapore    
Other Details:
Languages:  eng     Pagination:  931-4     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved.
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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