BioGenomics™ 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.
DescriptionTotal 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 65ºC for 10 minutes. The cDNA is in 1X RT buffer. (1X RT Buffer: 50mM Tris-CI, pH 8.3, 75mM KCI, 3mM MgCI2, 10mM DTT). The estimated cDNA concentration is about 2.5ng/ul. 1ul cDNA is sufficient for one PCR reaction.
Quality Control1. 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 (T5595-0010: 10 reactions).
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.
Unit:1ul cDNA is sufficient for one PCR reaction.
Donor Information:As reported
Storage Conditions:Store cDNAs at -20°C
Control PCR Condition:Ready First Strand cDNA:1ul10X PCR Buffer:2.5ul10mM dNTP:0.5ulControl Primers (5uM):1ulH2O, Nuclease-free:19.8ulTaq 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.
仕様
Size：10Tests
Source Antigen：Human
Grade：Molecular Biology Grade
Form：Supplied as a liquid in 1X RT Buffer: 50mM Tris-Cl, pH 8.3, 75mM KCl, 3mM MgCI2, 10mM DTT.
