RNA is extracted by using Qiagen RNeasy kit. 2 ug total RNA is then reverse-transcribed using an Applied Biosystems’ High-Capacity cDNA Reverse Transcription kit. 1 ul cDNA is sufficient for one PCR reaction. HAdCC cDNA is for research use only. It is not approved for human or animal use, or for application in in vitro diagnostic procedures.
The A260/A280 ratio of the cDNA is measured to be between 1.7-1.9 using a Beckman Coulter DU 800 spectrophotometer.
The cDNA is performance-tested in a control PCR using β-actin primers.