Greetings to all!!
DNA is isolated from infected cotton leaf.
The image is attached. It looks kind of shearing.
What are possibilities to use it for PCR?
It does look like partially degraded dna but it covers a wide range of large sizes so as PCR only amplifies short dna templates it should amplify ver well using these dna samples because the degradation is random and many larger fragments will contain your dna template sequence
DNA is extensively degraded, doing PCR with this DNA depends of the length of your amplicon, i.e., small amplicons will come whereas large amplicons won't
Hi Dr. Bawankar,
The DNA seems to be smearing significantly, but some remain visible and concentrated near the wells. However, be cautious as overloading the wells with large amounts of DNA can potentially result in smearing. It would help if you considered increasing the agarose percentage. Other factors contributing to smearing, are excessive gel running buffer (more than 5 mm above the gel surface, elevated voltage, and using old buffers, all of which can affect DNA movement on the gel.
Additional RNAse treatment may not be necessary as RNA is unlikely to interfere significantly due to the PCR's staging and cycling conditions. During the denaturation phase of PCR, any remaining RNA will be eliminated from the reaction.
Kind Regards,
Dharmesh
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