Magnetic quantitative reverse transcription PCR: A high-throughput method for mRNA extraction and quantitative reverse transcription PCR

Magnetic quantitative reverse transcription PCR: a high-throughput method for mRNA extraction and quantitative reverse transcription PCR.

Over the past few years high-throughput platforms for real-time quantitative PCR have become widely available. The cost of RNA extraction from a large number of samples are, however, quite notable. One method that stands out with respect to free up- or downscaling of sample size and reliability is the isolation of mRNA using oligodeoxythymidylate [oligo(dT)25]-coated magnetic particles. In comb...

High-throughput real-time quantitative reverse transcription PCR.

Extensive detail on the application of the real-time quantitative polymerase chain reaction (QPCR) for the analysis of gene expression is provided in this unit. The protocols are designed for high-throughput, 384-well-format instruments, such as the Applied Biosystems 7900HT, but may be modified to suit any real-time PCR instrument. QPCR primer and probe design and validation are discussed, and...

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

Design and optimization of reverse-transcription quantitative PCR experiments.

BACKGROUND Quantitative PCR (qPCR) is a valuable technique for accurately and reliably profiling and quantifying gene expression. Typically, samples obtained from the organism of study have to be processed via several preparative steps before qPCR. METHOD We estimated the errors of sample withdrawal and extraction, reverse transcription (RT), and qPCR that are introduced into measurements of ...

Quantitative reverse transcription-PCR comparison of tumor cell enrichment methods.

platelet plots are not always seen in cryoglobulinemia, although other interferences are usually detected with high efficiency. For comparison, whereas the Technicon H*3 appeared to provide a more immediate visual indication of sample cryoglobulins, large platelet count overestimates were still obtained in most cases. Examination of graphic plots is not used in all laboratories, and its efficie...