I have isolated DNA from animal tissue. The concentration of DNA has been measured as 200ng/microL. I am afraid that the genomic DNA might have contamination with RNA. Now, If do like to remove that RNA. How can I do that?
Have u done purification of DNA by Treating with RNAse? Its advised that you can do repurification of DNA even if you have done the previous.all d best
Be aware that RNAase preparations (even those commercial) can be contaminated with minute amounts of DNAase activity that upon long incubations could nick (or even ds-cut) your DNA sample. You can buy a DNAase-free RNAase, or just boil the RNAase enzyme (i.e. 5 min at 90-100ºC) to get rid of the DNAase activity (the RNAase will survive the treatment, see the Sambrook [Maniatis] manual for more details).
If you are isolating DNA manually, then just treat your DNA with RNAase. And if you are using DNA isolation+Purification kit, then don't worry about that, RNAase are included in the kit normally (Check the buffer composition).
you are advised to use RNAase.The DNA can be located by staining with low concentration of fluorescent interclating dyes such as EtBr or SYBR gold.
Dear Islam
For a typical prep I usually use 10 units of DNAse free RNAse . You may loose some DNA if you want to treat with RNAse after DNA Isolation hence it is always advisable to treat with RNAse-A & Protinsae-K while isolating itself. You can add 10-15 uL and incubate at 37C for 30 minutes or little longer no problem. If you want to denature you can incubate it at 80C for 30 min.
Best Regards
During genomic DNA isolation there are some steps which are very critical that can leads to RNA contamination,i used to get RNA contamination. Go for RNase treatment.
NEB says that their RNAse H digest 1 nmol RNA in 20 min at 37°C.
If you have large amounts of large RNAs, you may also consider a preliminary ethanol precipitation with lithium chloride ((0.2 volumes of an 8 M LiCl solution) before RNase treatment.
yes
Simply treat your isolated DNA with RNAase to get rid of RNA contamination. all the best
To remove genomic DNA (gDNA) from the RNA solutes, it treates with 10x reaction buffer with MgCl2 and RNase-free DNase-I diluted in water and incubate for 30 min at 37◦C. Ad 50 mM EDTA and incubate for 10 min at 65◦C
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