[Construction and identification of cDNA library from peripheral blood lymphocytes of Tibetans in Qinghai-Tibet plateau].
OBJECTIVE
To construct the full length cDNA library of the peripheral blood lymphocytes in Tibetans and to identify the quality of the library.


METHODS
The lymphocytes were separated from healthy Tibetan fresh blood, and total RNA was extracted. The full length cDNA library was constructed by SMART (switching mechanism at 5' end of RNA transcript) technique. The unamplified library was titered, and the percentage of recombinant clones were determined. The library was amplified, and the size of the inserts was identified by PCR.


RESULTS
The titer of the constructed cDNA library was 1.8x10(9) pfu/L with the percentage of recombinant clones above 98%, and that of the amplified library was 8x10(12) pfu/L. The insert's size ranged from 0.75 to 2.0 kb with the average length of 1.35 kb.


CONCLUSIONS
The constructed cDNA library of peripheral blood lymphocytes of Tibetans has excellent quality, and it lays a solid founclation for further screening and cloning hypoxia-related genes of Tibetans living at high altitudes.
Chen
Yun-tian
YT
Research Center for High Altitude Medicine of Qinghai University, Xining 810001, China. woshiyuntian@163.com
Wu
Jian-fang
JF
Cao
Yue
Y
Ge
Ri-li
RL
D044466:Asian People / Q000235:genetics; D002681:China / Q000208:ethnology; D004274:DNA, Recombinant / Q000235:genetics; D015723:Gene Library; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000502:physiology; D008297:Male; D008913:Minority Groups; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D018609:Tibet / Q000208:ethnology; D055815:Young Adult
D004274:DNA, Recombinant