发明名称 |
IMPROVED LYSIS AND REVERSE TRANSCRIPTION FOR MRNA QUANTIFICATION |
摘要 |
The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M guanidine thiocyanate, b) diluting the sample to an extend such that guanidine thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time. |
申请公布号 |
EP2147120(A1) |
申请公布日期 |
2010.01.27 |
申请号 |
EP20080749214 |
申请日期 |
2008.04.29 |
申请人 |
ROCHE DIAGNOSTICS GMBH;F. HOFFMANN-LA ROCHE AG;TATAA BIOCENTER AB |
发明人 |
BENGTSSON, MARTIN;KUBISTA, MICHAEL;STAHLBERG, ANDERS;STROEMBOM, LINDA |
分类号 |
C12Q1/68 |
主分类号 |
C12Q1/68 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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