Construction of cDNA library from the induced human peripheral blood lymphocytes(Hu-pBL) = 인체 말단 혈류 중의 임파구로부터 cDNA 군의 조립에 관한 연구

Lymphocytes were isolated from human periperal blood by Ficollpaque and immune interferon was induced in isolated lymphocytes in culture by combined treatment with a phorbol ester (12-o-tetracanoylphorbol 13-acetate, TPA) and Concanavalin A. Total RNA was isolated from mononuclear cells induced with TPA and Con A, and passed through oligo-(dT)-cellulose column to obtain mRNA. Poly(A)-containing RNA was transcribed into cDNA by AMV reverse transcriptase under appropriate conditions which increase the yield of full length cDNA. After alkaline hydrolysis of mRNA, the second cDNA strands were synthesized with hairpin loop as a primer. The loop of the ds-cDNA was specifically cleaved under appropriate conditions. cDNAs larger than 700 bp were purified on a preparative agarose gel. After a tailing step with dCTP, cDNAs were annealed to plasmid DNA tailed with dGTP. Recombinant plasmid DNAs were transformed in $\underline{E}. \underline{coil}$ LE392.
Advisors
Han, Moon-Hi한문희
Publisher
한국과학기술원
Issue Date
1985
Identifier
64478/325007 / 000831501
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생물공학과, 1985.2, [ [v], 60 p. ]

URI
http://hdl.handle.net/10203/28188
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=64478&flag=t
Appears in Collection
BS-Theses_Master(석사논문)
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