Removal of contaminating genomic DNA is an important step in many RNA based applications including RT-PCR. It  is also a necessary step to avoid false positive results. A frequent cause of concern among investigators performing quantitative RT-PCR is the occurence of false positives that is caused by genomic DNA contamination of RNA preparations. Because PCR is such a sensitive technique, a single copy of a gene can, theoretically be amplified and detected.

Generally DNase I is used to remove contaminating DNA. DNase is an endonuclease that cleaves DNA by breaking phosphodiester bonds.However, the DNAse has to be removed or inactivated from the RNA sample prior to RT-PCR.  There are different ways to achieve this

• Chelation with 20 mM EDTA - removes the Mg ions necessary for the activity of DNAse

• Heating at 70°C for 5 minutes - Inactivates the DNAse

• Proteinase K digestion followed by phenol/chloroform extraction and NH4OAc/EtOH precipitation - This extraction specifically cleaves the DNase I and precipitates the DNA, while the RNA remains in the aqueous phase and can be recovered by a subsequent precipitation.

Keeping your needs in mind we choose the protocol that's right for you.

Deliverables:

• Agarose gel picture of the RNA samples.
• Certificate of proof that it is free of genomic DNA.

Pricing: Starts from $200.00 and up. For additional information or for a quote This email address is being protected from spambots. You need JavaScript enabled to view it..