Tissue cDNA, First Strand, Mouse Adult Normal, Adipose, BioGenomics(TM)

Produktübersicht

• Artikelname: Tissue cDNA, First Strand, Mouse Adult Normal, Adipose, BioGenomics(TM)
• Artikelnummer: USB-T5595-0659
• Hersteller Artikelnummer: T5595-0659
• Alternativnummer: USB-T5595-0659-40
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

BioGenomics(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. 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 65C for 10 minutes. 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 spectro- photometer. A260/280 is between 1.8-2.0 (detected in 10mM Tris-Cl, pH 7.5). The ratio of 28S/18S is 1. 2. The RNA used for cDNA synthesis is treated by DNase I. It is tested as DNA free RNA by PCR. 3. The synthesized cDNA was 5 selected to ensure its full length. The cDNA was used as template for PCR amplification of b-actin gene. A 838 bp b-actin band was visualized on 2% agarose gel. 4. b-actin control primer pair is included in each package. It is enough for 10 PCR reactions. Control PCR condition: Reaction Mixture: PCR Ready First Strand cDNA: 1ul 10 x PCR Buffer: 2.5ul 2.5 mM dNTP: 4ul Control primers (25uM): 1ul H2O, Nuclease-free: 16.3ul Taq Polymerase(5u/ul): 0.2ul The PCR thermocycling: 94C x 2 minutes, 1 cycle 94C x 30 seconds, 55C x 45 seconds, 72C x 30 seconds, 35 cycles 72C x 5 minutes, 1 cycle. Then hold at 4C. Storage and Stability: May be stored at 4C for short-term only. For long-term storage, aliquot and store at -20C. Aliquots are stable for at least 6 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.