Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 1 de 1
Filter
Add filters








Language
Year range
1.
Asian Pacific Journal of Tropical Medicine ; (12): 931-934, 2012.
Article in English | WPRIM | ID: wpr-820592

ABSTRACT

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.


Subject(s)
Humans , DNA, Protozoan , Chemistry , Genetics , Gene Library , Giardia lamblia , Chemistry , Genetics , Giardiasis , Parasitology , Polymerase Chain Reaction , Methods , RNA , Chemistry , Genetics , Trophozoites , Chemistry
SELECTION OF CITATIONS
SEARCH DETAIL