Hi, I am a little confused working with total RNA extracted by the simple Trizol method, to avoid the genomic DNA contamination I want to treat it with DNAse 1 which is a Lyophilized powdered from Sigma cat # (D4263)https://www.Sigma-Aldrich.com/PK/en/product/sigma/d4263?gclid=Cj0KCQjw_fiLBhDOARIsAF4khR0ZHsByvb06kyqVx2PducHgQRPHPS1Y2KQ3eFKMgLvCPokVmoDKEJUaAq8PEALw_wcB. I have looked into the details on the website but couldn't found it helpful. Can anyone help me with its protocol for resuspension in water? also please let me know,
- Can I use water?
- how much water should I add?
- what will be my final concentration per microlitre (as I need 1unit for 1000ng of RNA.
It will be highly appreciated if someone can share their personal experience.
Thanking you in advance.
From the product page on the Sigma-Aldrich website:
The enzyme powder may be reconstituted in water or any buffer at pH 4.0-9.0, except phosphate buffer. Calcium chelators should be avoided. 10 mg/mL solution of DNAse I in 0.15 M NaCl may lose
Hi Laiba Shah,
You can use water to reconstitute your DNAse by adding 2ml (H2O) to the powder vial to have 2000U/2ml (=1U/µl). Keep your DNAse suspension in aliquots at -20°C to preserve its activity.
- Badges
- Science method