Quantification of RT-PCR Products: Ethidium Bromide—Stained Gel Analysis Compared With Fluorescent Detection Using an Automated Sequencer

Analysis of differential display RT-PCR products using fluorescent primers and GENESCAN software.

Differential display reverse transcription PCR (DDRT-PCR) is a procedure used to identify the induction or repression of gene expression. In most DDRT-PCR protocols, radioisotopes are incorporated during PCR and the cDNA products are detected by autoradiography. This report describes the fluorescent labeling of cDNAs and their detection on automated sequencers from PE Applied Biosystems. A fluo...

An integrated microsatellite length analysis using an automated fluorescent DNA sequencer.

Analyzing microsatellite instability (MI) in malignant tumors is thought to be useful for screening cancer patients to identify those patients with a higher risk of developing second malignant tumors. In this paper, we report a new, accurate, and efficient method of detecting MI using an automated fluorescent DNA sequencer and a computer that automatically calculates the size, height, and area ...

Use of the fluorescent dye PicoGreen for quantification of PCR products after agarose gel electrophoresis.

Exact quantification of the amount of amplified polymerase chain reaction (PCR) product is a crucial element in quantitative PCR approaches. Normally such quantification is based on the application of radioactive(2) or fluorochrome-labeled primers (4), HPLC (3) or densitometric-scanning of an agarose gel after staining with dyes like ethidium bromide (1). These quantification techniques often r...

Rapid quantitation of mRNA species in ethidium bromide-stained gels of competitive RT-PCR products.

A rapid method for quantifying the low-abundant mRNAs of the low density lipoprotein receptor and the 3-hydroxy-3-methylglutaryl coenzyme A reductase by competitive polymerase chain reaction is presented. This approach requires neither special labeling nor blotting procedures. For each analysis, a defined amount of total cellular RNA is co-reverse transcribed and co-amplified with a titration s...

Gel Purification of PCR Products. Fatemeh Mehrpooyan*