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Construction and screening of suppression subtractive hybridization library of renal cell carcinoma / 中华外科杂志
Chinese Journal of Surgery ; (12): 90-92, 2003.
Article in Chinese | WPRIM | ID: wpr-257723
ABSTRACT
<p><b>OBJECTIVE</b>To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).</p><p><b>METHODS</b>Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.</p><p><b>RESULTS</b>The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.</p><p><b>CONCLUSION</b>The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Gene Library / Adenocarcinoma, Clear Cell / Cell Line, Tumor / Genetics / Kidney Neoplasms / Methods / Nucleic Acid Hybridization Type of study: Diagnostic study / Screening study Limits: Humans Language: Chinese Journal: Chinese Journal of Surgery Year: 2003 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Gene Library / Adenocarcinoma, Clear Cell / Cell Line, Tumor / Genetics / Kidney Neoplasms / Methods / Nucleic Acid Hybridization Type of study: Diagnostic study / Screening study Limits: Humans Language: Chinese Journal: Chinese Journal of Surgery Year: 2003 Type: Article