Molecular Processing

Jean Dolce and Jian Tan
Molecular Processing Laboratory Pathology Core Facility Basic Science Building Rm S616 Email: Jean.Dolce@RoswellPark.org Jian.Tan@RoswellPark.org Tel: (716) 845-4420

LISTING OF FROZEN TISSUE RNA/DNA SAMPLES

Molecular Processing supports clinical scientific research throughout the Institute, contributing to the larger research efforts of independent investigators

DNA Frozen:  Genomic DNA is isolated using a DNA Purification Kit. DNA is purified from tissue by lysing of cells with an anionic detergent. Contaminants are removed using RNase and salt precipitation. Genomic DNA is then recovered by alcohol precipitation.  QC: 260/280 ratios are examined to confirm preparation purity and a DNA is run on an agarose gel to confirm presence of high molecular weight product.

RNA Frozen:  RNA is isolated using Guanidinium thiocyanate-phenol-chloroform extraction. Cells are homogenized using a reagent. RNA is then precipitated from the aqueous phase using isopropanol. QC: 260/280 ratios are examined to confirm preparation purity and an RNA aliquot is run on a bioanalyzer to confirm RNA integrity by generating an RNA Integrity Number (RIN) value. Specimens with a RIN of 7 or greater are generally suitable for expression analysis while those with a value of 5 to 7 yield less than optimal results for array studies. Specimens with a RIN less than 5 are not suitable for array studies, but may yield results for RT-PCR.