Microarray validation: factors influencing correlation between oligonucleotide microarrays and real-time PCR

An oligonucleotide microarray for multiplex real-time PCR identification of HIV-1, HBV, and HCV.

We describe a novel microarray-based approach for simultaneous identification and quantification of human immunodeficiency virus type 1 (HIV-1) and hepatitis B and C viruses (HBV and HCV) in donor plasma specimens. The method is based on multiplex real-time RT-PCR performed within the microarray hydrogel pads. Double-stranded amplification products are simultaneously detected using nonspecific ...

Factors Influencing Real-Time RT-PCR Results: Application of Real-Time RT-PCR for the Detection of Leukemia Translocations

A variety of factors that influence real-time reverse transcriptase (RT)-PCR results were examined. Realtime reaction volumes can be decreased from the manufacturers suggested volumes (50μl) to as little as 5μl. Errors introduced through volume handlings can be significantly mitigated through the use of electronic pipettes. Random hexamers are compared against the use of a specific reverse tran...

Critical Factors for Successful Real-Time PCR

Validating Microarray Data Using RT Real-Time PCR

This paper describes the use of real-time PCR for the confirmation of microarray data. Current publication guidelines require that all microarray results are confirmed by an independent gene expression profiling method. Real-time PCR is the method of choice for most researchers but it is not without drawbacks. The first step in confirming array results by real-time PCR is selection of gene-spec...

Critical Factors for Successful Real-Time PCR