First-Strand cDNA is synthesized from RNA isolated from a wide variety of documented human normal tissues, human diseased and tumor tissues, mouse, rat, monkey, dog, and plant tissues. Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques. 10 µg total RNA was primed by an oligo dT primer and reverse transcription (RT) using MMLV reverse transcriptase in 40 µl final volume. RT reaction was stopped by heating at 65°C for 10 minutes. The cDNA is delivered in 1x RT buffer (1x RT Buffer: 50 mM Tris-Cl, pH 8.3, 75 mM KCl, 3 mM MgCl2, 10 mM DTT). 1 µl cDNA is sufficient for one PCR reaction. The 5' end of human clathrin cDNA (a 6 kb gene) has been amplified by PCR from all of these cDNAs.Features • Ready-to-use for PCR • Oligo dT primer used to ensure the presence of the entire 3' end of cDNA • 12 kb PCR amplicon has been successfully obtained with some templates indicating intact cDNAs • The largest selection of cDNAs from different tissues on the marketApplications • Immediate PCR amplification of known genes • Verification of genetic mutation • Comparison of a specific gene between different tissues • Analysis of mRNA alternative splicing • Gene cloning and target sequencingQuality Control • The integrity of the RNA used for cDNA synthesis is examined by visual inspection for the presence of intact bands of 18s and 28s ribosomal RNA when electrophoreses on a denaturing agarose gel. • The quality and purity of total RNA were tested by spectrophotometer. A260/280 is between 1.8 and 2.0 (detected in 10 mM Tris-Cl, pH 7.5). The ratio of 28S/18S is =1. • The RNA used for cDNA synthesis is treated by DNase I, and is tested as DNA-free RNA by PCR. • The synthesized human, animal, and cell line cDNA was selected to ensure its full length. The cDNA was used as template for PCR amplification of ß-actin gene and an 838 bp ß-actin band was visualized on 1% agarose gel. ß-actin control primer is included. It is enough for 10 PCR reactions. • The synthesized plant cDNA was used as template for PCR amplification of chloroplast gene. A 458 bp chloroplast band was visualized on 1% agarose gel. Chloroplast control primer is included. It is enough for 10 PCR reactions. For lot specific information such as Diagnosis and Donor Information, please refer to the Datasheet below.Important Notice: This tissue product is manufactured by a third party and is presented with the pathologic diagnosis supplied by the provider. Due to the variability inherent in pathology samples and the tissue cores derived from them, LSBio recommends that upon receipt, customers should review the histology/pathology of each specimen in order to confirm that it is consistent with the diagnosis supplied. Storage Conditions: Store at -20°C.Other lots of this item may be available upon request.