One-Step RT-PCR without Initial RNA Isolation Step for Laser-Microdissected Tissue Sample

NOTE 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 ...

One-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...

One-step RT-PCR for screening microdissected tissue samples.

The technology of laser-capture microdissection (LCM) enables the easy extraction of single cells or defined groups of cells from a tissue section (1,2). This technique uses a laser pulse to activate a plastic polymer, which binds to and captures cells from a frozen or paraffin-embedded tissue section. Captured cells present on this solid support can then be used for nucleic acid and/or protein...

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*

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...