Cryophile gDNA Removal Kit
The Cryophile gDNA Removal Kit is designed for removal of contaminating gDNA from RNA prior to reverse transcription. After DNase treatment the Cryophile gDNA removal Kit is easily inactivated by a moderate rise in temperature (58°C). gDNA removal and reverse transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.
The Cryophile gDNA removal Kit contains a final Mg concentration of 3 mM. As a result, carryover of magnesium to the RT reaction must be considered. If 10 μl of digested RNA is used, the magnesium concentration in a 20 μl RT reaction will increase by 1.5 mM. If the RT kit used for downstream cDNA synthesis is incompatible with additional magnesium, the following steps can be implemented:
• Dilute the DNase treated RNA prior to reverse transcription
• Use a smaller portion of the DNase treated RNA for reverse transcription
• Add EDTA to a final concentration of 3 mM prior to reverse transcription
DNase treatment can also be performed on entire RNA preps (up to 50 μl). It is worth considering that that Mg may be present in the remaining RNA, which might influence its long term stability. If the volume of RNA exceed 20 μl, it might be beneficial to double the amount of the provided DNase in the kit. Avoid preheating of decontaminated RNA and primers above 60°C prior to reverse transcription.
Source
Cryophile gDNA removal Kit is a mutated version of dsDNase from arctic shrimp. It is recombinantly expressed in a non-animal host (yeast).
Kit contents
gDNA removal Kit containing DNase(1 vial) and 10x Reaction Buffer (2 vials).
Sufficient for 50/ 250 reactions (depending on pack size)
Expiry
See label.
Storage
Store at -20°C. Aliquotation of the reaction buffer is recommended to avoid repeated freeze-thaw cycles.
Keep reaction buffer cold.
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