I have done RAPD Analysis for one plant with its Invivo tissue and in vitro tissue samples. I have used 31 primer sets, so now i dont getting how much maximum variation can be considered for this analysis. i mean mostly 80 to 100% polymorphism were seen in two or more different species of plant, but here in our case the two different samples of the same plants have been used, one is in vitro lief and another one is in vivo keaf sample. so anyone has any idea regarding this then please help me to analyse the data and the way to going for the final conclusion for this kind of study.
Hi!
Before going to answer I would like to know what made you to perform RAPD in two ways in vivo n in vitro any specific reason, quite interesting. I would like to check/see both gel images at least for 2, 3 primers. Do you see any variation from in vivo to in vitro!!!
Vast literature is available how to score/analyze RAPDs. First you have to develop/generate 0, 1 data matrix for presence and absence of bands/alleles then using that you can develop a tree to check the evolutionary relationship among the members (variety of software available). One advise, once you type check for spelling mistakes too. RG shows/recognize spelling mistakes (keaf). You can send a msg. to me if you do not want to disclose the reason.
Check the attached articles.
Earlier I answered many questions related to RAPD. If you have patience kindly check for answers. Good Luck!!!
Thank you jetty ramadevi,
I have to do the RAPD for invitro n in vivo to check the polymorphism produced in invitro sample produced from mother plant with help of tissue culture. Yes i got the polymorphism in 60% primers we ised.
That's fine.
Can I have a look at your RAPD profiles at least for one RAPD primer for both in vitro n in vivo then I advise better. Attach the gel profile with all details so that I come to know how they worked and guide you better.
here i attached gel picture showing band structure for some primer sets of invitro and invivo plant DNA samples.
Here in image a lane means in vitro and b lanes means invivo.
Hi!
I have seen your gel picture today that too after watching same question in Q.s wondering I advised him and gone to your old question. Now a days problem in receiving mails reg. RG msg.s. Better contact by sending personnel msg. then I come to know.
I told already some primers will not give many bands so you can put aside in the sense they do not work (good) for these samples. You note down the primers where you got many bands after check all primers take those and run the RAPD-PCR with all your samples and generate binary matrix (0,1) for all primers what ever you have used. Once it is done create dendrogram (many software available). I sent publications too, seems still you have not followed or you need more explanation, let me know.
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