I am trying to extract DNA from cannabis leaf samples, and at first had no problems, but now I can't get any DNA out. In the last step of extraction I can't see anything in the tube. Any suggestions as to what the problem might be?
Are you doing the DNA isolation with a commercial kit? If not and you are using a protocol. Have you prepared any new reagent again? It sounds that the last extractions you did where done a while ago. If you prepared some reagent again maybe that can be the problem.
The other problem can be the tissue. Are you collecting and using the leaves to do the isolation in the same way? I don´t know maybe you collected and the preserved in the freezer. Some plants don't like that. I you are doing exactly the same, can be as Stalis suggest.
First let me What method you had been used and facing problem in where exactly,then I can suggest you precisely or else you can follow CTAB method thats very selective for DNA Extraction from Plant source.
you are using DNA extraction kit? i thing problem in lysis of cell wall. because plant cell wall is much harder than bacterial cell wall so u can check your lysis buffer or add more amount of lysis buffer.
CTAB is the best protocol in any way. And yes i agree with maimuthu that it seems that problem is in cell lysis. perfrom your cell lysis step efficiently. you will definitely get dna. Good luck
First of all grind the leaves properly.
CTAB method is good for isolation of DNA.
Check the pH of all the reagents used. pH of the reagents is major concern in DNA isolation.
precipitate the DNA carefully according to the protocol.
Try with the some one other's reagents.
May be helpfull...good luck
Please tell us what protocol you are using.
CTAB is a good one indeed.
Remember:
1. Even if your lysis/homogenization only opens up 20% of your cells,
2. Even if after your last step you can't see anything in the tube,
You probably still have lots of good DNA you can do fantastic PCRs with!
You don't know anything about your DNA untill you've redissolved a potentially invisible DNA pellet in H2O (and if you want, measured on a spectrofotometer/nanodrop or run on a gel), and you've used it as template for PCRs.
So if you give us a better problem description and what you've done so far, we can help better.
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