Cambio - Excellence in Molecular Biology

Enzymes for Molecular Biology

Enzymes for Molecular Biology: Heat labile & Salt Tolerant Enzymes

New range of enzymes for 2019. Our new range of enzymes include novel enzymes specifically designed or engineered to be amazingly tolerant to high salt concentrations, function at very low temperatures and heat inactivated.

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 enzyme 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.

Cambio

Catalogue No.DescriptionPack SizePriceQty
  • Change to:
  • £
CA-1728-250Cryophile gDNA Removal Kit250 reactions POA Quantity Add to Order
CA-1728-50Cryophile gDNA Removal Kit50 reactions POA Quantity Add to Order

Description

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.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

Set up the reaction as follows:

1. Add 1 μl from Cryophile gDNA removal enzyme to an empty tube on ice  
Note: Use the same tube as intended for reverse transcription.

2. Add 1 μl 10x Reaction Buffer for each 10 μl of RNA

3. Add 8-50 μl of your RNA
Note: If same-tube reverse transcription is intended, make sure that the total volume of the gDNA removal Kit reaction does not exceed the maximal input limit of the RT-kit.

4. Incubate at 37°C for 10 minutes (gDNA digestion)
Note: Incubation can also be performed at 25°C for 20 minutes.

5. Incubate at 58°C for 5 minutes (Salt-tolerant gDNA removal enzyme inactivation)

6. Proceed to reverse transcription.
Note: Add RT reagents of your choice directly to the Salt-tolerant gDNA removal Kit reaction mix.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200