I extracted RNA from the fresh tissues using RNA kit form GCC Biotech, normally during the Quality check using electrophoresis it showed Double band but today in my case i could get only one single thick band and when checked using Nanodrop i got concentration of about 400ng/ul with 260/280 value of 2.01 and 260/230 value of 1.5.
Please can some suggest me the possible reasons?
Hello, Amit Kumar Pradhan,
possible reasons are:
- too short electrophoresis time for bands separation (one thick band could be unresolved two bands)
- did you use some DNA/RNA ladder? If yes, your thick peak correspond to 28S or 18S RNA?
- your spectroscopic parameters of isolated total RNA are quite good/no problem I think
- maybe your isolated RNA is partially degraded due to ubiquitous enzymes RNAses, I would recommend you to follow this simple protocol for agarose gel electrophoresis, which protect RNA against RNAses (Article Bleach Gel: A Simple Agarose Gel for Analyzing RNA Quality
)Hope this help
Martin
Thank You Martin Bartas it is a good suggestion i will definitely try it.
Hello Amit,
Agreed with Martin that ur gel might ran for too short time and bands remain unresolved but i dont think their is any degradation as u can see a thick band without any smearing....
possible reason for single thick band might be rRNAs only...
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