منابع مشابه
RT-PCR without RNA isolation.
Reverse transcription-PCR (RT-PCR) has traditionally required time-consuming RNA extraction and purification. This report demonstrates that one can completely avoid the RNA extraction step in RT-PCR by basing the comparison of samples on cell number rather than micrograms of total RNA. A new method for lysing cells while preserving RNA is described. RT-PCR is carried out (i) by rapidly freezing...
متن کاملNOTE Pathology One-Step RT-PCR without Initial RNA Isolation Step for Laser-Microdissected Tissue Sample
One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RTPCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis ...
متن کاملOne-step RT-PCR without initial RNA isolation sStep for laser-microdissected tissue sample.
One-step RT-PCR procedure without initial RNA extraction step is tested for laser microdissected tissue sample. Unfixed cryosections of liver and kidney tissue of male SD rats were cut using laser microdissection system and directly used as templates for RT-PCR study. To check the sensitivity, 5, 25, 125, and 625 hepatocytes were cut and put in PCR-tube. After DNase treatment and cDNA synthesis...
متن کاملCompetitive RT-PCR: Preparation of Competitor RNA.
INTRODUCTION The first step in competitive RT-PCR is the synthesis and purification of the synthetic competitor. This is an RNA molecule designed to be reverse-transcribed and PCR-amplified with the same efficiency as the endogenous transcript of interest. Once the competitor molecule has been prepared, as described in this protocol, competitive PCR can be carried out, as described in Comptetit...
متن کاملSimultaneous quantification of four RNA targets by multiplex, real-time RT-PCR without optimization.
Researchers carrying out quantitative, multiplex, realtime PCR and RT-PCR require master mixes that provide accurate, simultaneous quantification of multiple targets in the same reaction without time-consuming optimization. Our results at Wyeth Research show that QuantiTect® Multiplex Kits from QIAGEN are well suited for multiplex analysis of up to four RNA targets in two-step and onestep real-...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1996
ISSN: 0736-6205,1940-9818
DOI: 10.2144/96216rr02