title: Construction and characterization of a full-lengh cDNA library from non-fresh Giardia lamblia.
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.
authors.affiliation: School of Laboratory Medicine, Jilin Medical College, Jilin 132013, PR China.
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Guo
authors.forename: Jie
authors.initials: J
authors.lastname: Jiang
authors.forename: Wen-Yu
authors.initials: WY
authors.lastname: Zheng
authors.forename: Ming-Luan
authors.initials: ML
authors.lastname: Li
authors.forename: Xi-Feng
authors.initials: XF
authors.lastname: Tian
authors.forename: Xian-Min
authors.initials: XM
authors.lastname: Feng
authors.forename: Yue-Hua
authors.initials: YH
authors.lastname: Wang
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Ju
authors.forename: Yue-Qiong
authors.initials: YQ
authors.lastname: Kong
mesh_terms: D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D015723:Gene Library; D016829:Giardia lamblia / Q000737:chemistry / Q000235:genetics*; D005873:Giardiasis / Q000469:parasitology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D053058:Trophozoites / Q000737:chemistry
chemical_list: D016054:DNA, Protozoan; D012313:RNA