What You Need To Provide
Between 50ng and 100ng of total RNA but we can work with as little as 5ng of good quality RNA.
What Genome Explorations Provides RNA Quality Check and Quantitation
Each RNA sample is quantitated using a dual beam spectrophotometer and checked for integrity (QC) using an Agilent Bioanalyzer 2100 "Lab on a Chip" system. Investigators are notified immediately if the RNA is degraded or otherwise unsuitable for GeneChip application.
Double Round Amplification and Biotin Labeling
Samples that pass our QC criteria are processed to synthesize First and Second Strand cDNA. This double stranded cDNA is used in an in vitro transcription step to yield unlabeled cRNA. More double stranded cDNA is generated through another round of first and second strand synthesis. Another in vitro transcription step synthesizes amplified biotin labeled cRNA. (Optional: The cRNA is quality checked and upon client request, can be first hybridized to an Affymetrix Test 3 chip to check for transcript integrity. Transcript quality is assessed by comparing the 5':3' ratios of a collection of house keeping genes). Again clients will be notified immediately if the transcript quality in unsuitable for the GeneChip application.
GeneChip Microarray hybridization and data analysis
Samples that pass all our QC criteria will be hybridized to the requested GeneChip arrays. Microarray data is supplied to Investigators as comprehensive lists of genes that are significantly regulated in TEST Vs REFERENCE samples. All unfiltered, complete data sets are also provided.
