Catalog Number ASD-9038
Quantity 20 reactions
Description Applied StemCell’s cDNA is synthesized from a highly pure and intact total RNA that was isolated via a modified guanidine thiocyanate technique and devoid of genomic DNA contamination. Five ug of total RNA was reverse transcribed into a single strand cDNA using oligo–dT primer in 20 mL total volume. RT reaction was stopped by heating at 85°C for 5 minutes. Using 1 µl of cDNA per PCR reaction is sufficient for 20 typical PCRs. Applied StemCell’s cDNA can be used in gene expression and cloning studies, analysis of mRNA alternative splicing, gene mutation analysis and other molecular biology fields.
Fibroblasts are isolated from an individual who has been clinically diagnosed with Mucopolysaccharidoses. Mucopolysaccharidoses are a group of metabolic disorders caused by the absence or malfunctioning of lysosomal enzymes needed to break down molecules called glycosaminoglycans – long chains of sugar carbohydrates in each of our cells that help build bone, cartilage, tendons, corneas, skin and connective tissue.
Origin Human, single donor
Development Period Postnatal
Disease State Mucopolysaccharidoses (MPS)
Test Conditions Quality of RNA used in cDNA synthesis is determined visually, analyzing 18S and 28S ribosomal bands by non-denaturing agarose gel electrophoresis. The RNA purity level is determined by spectrophotometry to obtain the A260/A280 ratio, which must range from 1.8-2.1. The RNA used in cDNA synthesis is treated with RNase-free DNase to remove any genomic DNA and is tested as DNA free by RT-PCR. The cDNA functionality is assayed by RT-PCR using primers for housekeeping gene GAPDH.