Tissue cDNA, First Strand, Plant Normal , Plant, Arabidopsis, BioGenomics™
|Molecular Biology||Storage: -20°CShipping: Dry Ice|
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.
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 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.
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
Store cDNAs at -20°C
Form: Supplied as a liquid in 1X RT Buffer: 50mM Tris-Cl, pH 8.3, 75mM KCl, 3mM MgCI2, 10mM DTT.
Unit: 1ul cDNA is good for one PCR reaction.
Plant Name: Arabidopsis
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.
Control PCR condition:
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:
94°C x 2 minutes
94°C x 30 seconds, 55°C x 45 seconds, 72°C x 30 seconds, 35 cycles
72°C x 5 minutes.
Then hold at 4°C.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.