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

Product Overview

• Article Name: Tissue cDNA, First Strand, Human Adult Normal, Brain, BioGenomics(TM)
• Biozol Catalog Number: USB-T5595-0025
• Supplier Catalog Number: T5595-0025
• Alternative Catalog Number: USB-T5595-0025-40
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

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. 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 65C 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). 1ul cDNA is good for one PCR reaction. The 5 end of human clathrin cDNA (a 6kb gene) has been amplified by PCR from all of the cDNAs. 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 The largest selection of cDNAs from different tissues on the market Documentation of tissues clinical histories available (additional cost) 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 Storage Conditions: Store cDNAs at -20C Control PCR Condition: Ready First Strand cDNA: 1ul 10X PCR Buffer: 2.5ul 2.5mM dNTP: 0.5ul Control Primers (5uM): 1ul H2O, Nuclease-free: 19.8ul Taq Polymerase (5u/ul): 0.2ul PCR Thermocycling: 94C x 2 minutes, 1 cycle. 94C x 30 seconds, 55C x 30 seconds, 72C x 30 seconds, 35 cycles. 72C x 5 minutes, 1 cycle. Then hold at 4C. Unique Features of cDNA: PCR Ready First Strand cDNA is made from our high-quality total RNA. Each cDNA is a premade, tissue-specific ''pool of first strand cDNA from which full-length genes can be amplified by using sets of gene-specific primers. They can also be used to study tissue-specific gene expression and to find not only polymorphic forms of mRNA but also mRNA belonging to a multigene family. The following features have been integrated into our cDNA products to ensure their superior quality. 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 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, and 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 and an 838bp b-actin band was visualized on 1% agarose gel. b-Actin control primer is included (T5595-0010: 10 reactions). 1. Three formats of cDNA products are available There are three types of cDNA products to choose from: PCR ready first strand cDNA, tumor/adjacent normal paired cDNA, and cDNA panels. 2. RNAs with high quality are used for cDNA synthesis The cDNA synthesis will not be successful using degraded or contaminated RNA, because degraded RNA will not generate full-length cDNA, and contamination may inhibit the synthesis of cDNA. The RNAs isolated by BioChains proprietary technology are pure and intact from primary tissues. These RNAs therefore ensure a very high quality of cD