I am extracting DNA from the leaf samples of T. arjuna, My problem is that if i reduce the washing steps then protein impurities remains in DNA while i increase the washing steps, then DNA is getting in very low quantity. Please suggest me how can i overcome this problem?
It is not clear from your answer which protocol you are using. I can try helping if you tell me the protocol or the reference.
Mam I am using modified CTAB method of Vishal P. Deshmukh ( Sant Gadge Baba, Amravati university). Before i extracted and my purity index was around 1.5, but presently i am in NIT, i tried many times here but not became successful, here my purity is 1.1 and 1.2 , checked my nanodrop method.
I went through the protocol. Can you please specify what is your incubation time in isopropanol and ethanol since they are not indicated in the protocol? You can try increasing the incubation times for both. Also, there are many washing steps in the protocol. So, the yield is bound to be less. Try to dissolve your DNA in less volume of TE buffer or dd water, anything you are using, in the final step.
I incubate for 30 min on 70 degree in water bath after adding the resuspension buffer and 10%SDS. after that place the sample on ice for 1 hours.
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