I did DNA purification using nucleospin purification kit to my paraffin embedded tissue to detect EBV Xhol site by Xhol restriction enzyme but when i did restriction enzyme cutting and run them on gel (4%), there is no band for all my samples.
I need your suggestion in this case to get obvious band after purification?
Did you quantify your DNA, or visualized on gel before any digestion ?
Hi abhijeet
Yes , i quantified them already (Conc. about 25 ng/ul) this is the lowest one and i visualized them before restriction assay (RE) cutting
All my samples have bands but after i did DNA purification of excised gel bands using Nucleospin kit and doing RE cutting procedure, all my bands did not appear.
What is your suggestion?
Thanks
Many times the gel extraction do not work and you will not get much DNA after gel purification. Thus, quantifying the DNA after gel extraction must also be performed and restriction enzyme volume should be optimized according to the DNA concentration. Use several replication of the DNA in gel and pool the DNA extracted after gel purification to good higher concentration.
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