TaqMan Array Cards in pharmaceutical research.
TaqMan Array Cards are high-throughput, accurate, sensitive, and simple-to-use tools for quantitative analysis of mRNA or miRNA transcripts using a real-time PCR protocol. They utilize a microfluidic card with 384 reaction chambers and eight sample loading ports. For studies of coding transcripts, the reaction chambers are preloaded with user selected or predefined panels of Applied Biosystems TaqMan Gene Expression Assays. These assays enable real-time monitoring of a PCR reaction via hydrolysis of an oligonucleotide probe which has been dual labeled with fluorescent dye and quencher. Applications of TaqMan Array Cards include verification and follow on testing of microarray results, as well as hypothesis driven testing of panels of genes selected for their biological functions and relationships. This chapter describes a protocol for assaying transcription in cultured cells using methods optimized to minimize hands-on time and pipetting steps by skipping RNA isolation and generating cDNA directly in Ambion Cells-to-C(T) lysis solution.
Keys
David N
DN
Molecular Biology Division, Life Technologies Corporation, Foster City, CA, USA. david.keys@lifetech.com
Au-Young
Janice K
JK
Fekete
Richard A
RA
D000818:Animals; D002457:Cell Extracts / Q000737:chemistry; D002478:Cells, Cultured; D004345:Drug Industry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D016133:Polymerase Chain Reaction; D051381:Rats; D048348:Reverse Transcription; D019914:Taq Polymerase / Q000378:metabolism*; D013997:Time Factors
D002457:Cell Extracts; D019914:Taq Polymerase