Tissue cDNA, First Strand, Rat Adult Normal, Brain, BioGenomics(TM)

BioSelect(TM) Tissue cDNA: cDNA is supplied as First Strand, Multiple Tissue Panels, and Matched Pairs. PCR-ready First Strand cDNA is tissue specific and are ready-to-use for gene discovery or expression analysis. Over 350 cDNAs from human adult and fetal normal tissues, human diseased and tumor tissues, rat, mouse, monkey and plant tissues are included in this extensive collection. PCR Ready First Strand cDNAs is an excellent source of tissue specific, PCR-ready cDNA, and it can be immediately used for gene discovery or expression analysis. First-Strand cDNA is synthesized from RNA isolated from a wide variety of documented human adult and fetal normal tissues, human diseased and tumor tissues, mouse, rat, monkey and plant tissues. Total RNA used for cDNA synthesis is isolated by modified guanidine thiocyanate techniques. 11ug total RNA was primed by an oligo dT primer and reverse transcribed by MMLV reverse transcriptase in 40ul final volume. RT Reaction stopped by heating at 65 degrees C for 10 minutes. The cDNA is in 1X RT buffer. (1X RT Buffer: 50 mM Tris-Cl, pH 8.3, 75mM KCl, 3mM MgCI2, 10mM DTT). Contents: 40ul per package. 1ul cDNA is good for one PCR reaction. Features: Ready to use for PCR Oligo dT primer used to ensure the entire 3' end of cDNA is present With some cDNA used as templates, 12kb PCR amplicon was obtained to ensure the intactness of cDNA Applications: 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 sequencing Quality Control: . 1. 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. 2. The RNA used for cDNA synthesis is treated by DNase I, and is tested as DNA free RNA by PCR. 3. The synthesized cDNA was 5