while isolating DNA from animal tissue I usually get bands in the upper region of the gel. But today after DNA isolation AGE was done at 100v for 30 minutes and the gel documentation yielded the following result. Iam confused about the bands seen on the gel. Whether they are RNA/Protein contamination in my DNA? or degraded DNA itself? the picture of the gel is attached along with.
To the 10ul samples, add ChIP elution buffer to a final volume of 50 uL
1. Add 1 uL Proteinase K and incubate at 58°C for 12 hours.
2. Add 1 uL of RNaseA (10 mg/mL, Sigma) and incubate for 60 minutes at 37°C.
3. Load 3 uL on a 1% agarose gel.
hope this will help you
Hi Keerthy,
First of all, Do I understand this is a gel of total DNA? Normally total DNA will appear distributed all along the gel , with great bands in the upper region... maybe this happen because the DNA has been degraded in small fragments. Are you sure about the quality of the sample? (defrost of the tissue...a new DNA isolating protocol...too much time at store...)
Best wishes
Dear David,
I have isolated total genomic DNA from snail tissue samples. The samples are preserved for about 3-12 months in ethanol. I have attempted a new DNA isolation protocol. Usually I was getting PCR products from DNA isolates. But for the last 3-4 months the DNA isolation from snail tissue fails. Do you thing that the bands on the gel is Protein or RNA contamination? expecting your reply.
I think those bands are not the interest of your products. That might be due to primer dimer which used to appear less that 100 bp..
Hi Keerthy,
I´m not sure about contamination...normally the result when you have contamination is the visualization of non expected bands in your gel... You said you are working with snails...are you using a DNA protocol specific for molluscs? Sometimes molluscs are problematic because have high levels of mucopolysaccharides on their tissues and can interfere with the DNA extraction. Moreover, when you observed the change in your results...you have changed the taxonomic group of the samples (even at species level..)
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