I have extracted the Genomic DNA from Candida albicans. Instead of a High size band i got a single very bright very below in gel. What it can be and why it happened?
Note: I did not use RNAse treatment.
Dear Sandeep,
Your results can be explained by the presence of nucleases. Do check for the same.
Abhishek
Dear Sandeep,
Presence of RNA is definitely the cause of such results. Add more RNA-ase and incubate a bit longer during the DNA extraction.
Mirsad
Something looks fundamentally wrong here. Yes, there is tRNA or possibly some other molecule binding the ethidium bromide. BUT there is not so much as a trace of a smear of gDNA in any of the lanes.
Fungi can be *very* resistant to lysis. Make sure that you are using a protocol specifically designed for Candida. Attached is an example.
Article Comparison of Six DNA Extraction Methods for Recovery of Fun...
RNase addition could be the best solution to get rid from this problem.
Thank you all for your valuable suggestions.
I have added RNAse and got Genomic DNA but the concentration is low (I think bcoz of Band intensity). I have used "Miniprep method for Genomic Dna isolation" from "Cold Spring harbor Laboratory Protocols".
Dear Kaushik
I got same DNA with you.
I speculated there are nucleases binding with DNA, so i added phenol to my DNA liquid to get rid of those proteins. Finally, i got pure genomic DNA showed normal band on agrose gel.
i attache my experiement steps here and hope it helpful to you.
1. Dissolve the DNA in 500ul TE.
2. Add 500ul phenol, mix well.
3. spin at 4500g, 20 min.
4. remove supernatant to a new EP tube, add 500ul 24:1 ,mix well, spin at 4500g, 20 min.
5. remove supernatant to a new EP tube, do the precipitation again.
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