Thermo Scientific
DNAse 1
164087
During cDNA synthesis, for the real time analysis, the RNA has to be very pure and should not contain any traces of DNA. During the conventional Trizol method of RNA purification, DNA will be precipitated along with the RNA . DNAse 1 kit can be used to degrade any trace of DNA in the sample, which is a very important aspect in cDNA conversion.
cDNA synthesis
Total RNA
After the RNA precipitation, the RNA is diluted in 1X DNAse buffer and 1 microliter of DNAse1 is added. Then it is incubated at room temperature for 15 minutes. After incubation, 1 microliter of stop solution is added and heat inactivated at 70 degrees for 10 minutes. Finally the RNA is dissolved in RNAse free water concentration is recorded
Do not prolong the heat inactivation step
Good quality RNA is obtained with out any traces of DNA
None
Simple procedure
No drawbacks
Very useful for people working on real time PCR and cloning