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You are here:Home » Cloning » Cloning-cDNA » Tissue cDNA, First Strand, Human Fetal Normal, Heart, BioGenomics™

Tissue cDNA, First Strand, Human Fetal Normal, Heart, BioGenomics™

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Specifications

Catalog #T5595-0381
BioSelect™ 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.
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°C for 10 minutes. The cDNA is in 1X RT buffer. 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 2% agarose gel. b-Actin control primer is included (T5595-010).
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
2.5ng/ul
Supplied as a liquid in 1X RT Buffer50 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. Contents
40ul or 10ul per package
Human, normal, fetal heart, 28 weeks old
Store cDNAs at -20°C
Ready First Strand cDNA1ul
10X PCR Buffer2.5ul
2.5mM dNTP4ul
Control Primers (25uM)1ul
H2O, Nuclease-free     16.3ul
Taq Polymerase (5u/ul)0.2ul
PCR Thermocycling
• 94°C x 2 minutes, 1 cycle.
• 94°C x 30 seconds, 55°C x 30 seconds, 72°C x 30 seconds, 35 cycles.
• 72°C x 5 minutes, 1 cycle. Then hold at 4°C.


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