United States


cDNA

Introduction to cDNA

Complementary DNA (cDNA) is DNA synthesized from a RNA template. Total RNA isolation is performed at our high performance facility by using modified guanidine thiocyanate techniques which ensure consistency. Our Total RNA sources originate from a wide variety of human tissues such as: human adult and fetal normal tissues, human diseased and tumor tissues, as well as, mouse, rat, monkey, and plant tissues.

After RNA isolation, first strand cDNA is synthesized with MMLV reverse transcriptase with low RNase H activity. BioChain’s cDNA can be directly use as a template for r polymerase chain reaction (PCR) or real-time PCR. It is also ideal for gene discovery or gene expression analysis.

We carry the largest selection of tissue cDNA in the research market (358 cDNAs) which includes human cDNA and tumor cDNA.

Features

  • Ready to use for PCR
  • Oligo dT primer used to ensure the presence of the entire 3' end of cDNA
  • 12 kb PCR amplicon had been successfully obtained with some templates indicating intact cDNAs
  • The largest selection of cDNAs from different tissues on the market
  • Documentation of tissues' clinical histories available
  • 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

  • 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 10 mM Tris-Cl, pH 7.5). The ratio of 28S/18S is ≥1.
  • The RNA used for cDNA synthesis is treated by DNase I, and is tested as DNA free RNA by PCR.
  • The synthesized human, animal, and cell line cDNA was selected to ensure its full length. The cDNA was used as template for PCR amplification of ß-actin gene and an 838 bp ß-actin band was visualized on 1% agarose gel. ß-actin control primer is included. It is enough for 10 PCR reactions.
  • The synthesized plant cDNA was used as template for PCR amplification of chloroplast gene. A 458 bp chloroplast band was visualized on 1% agarose gel. Chloroplast control primer is included. It is enough for 10 PCR reactions.
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