I want to prepare 1/2 MS supplemented with PEG, in % (1.5, 3.0, 4.5 and 6.0). I read in an article (J. Exp. Bot. (2000) 51 (350): 1555-1562) that equal volume (i.e., PEG volume= volume of 1/2 MS media in the solidified plate) of filter sterilized(PEG is unstable at high temperature) PEG should be poured over the solidified half MS plate. This plate should be kept for 24 hours (at what temperature the plate should be stored after pouring PEG?) for the PEG to diffuse into the half MS media.
Further, another paper (http://www.gantlet.org/protocols/pegp.pdf) says that to obtained required concentration of PEG in media, add twice the concentration of PEG.
Am I following the right protocol? Could any one help in smoothing the protocol based on what I wrote above. Your valuable comments are awaited!
Thanks a lot!
http://www.gantlet.org/protocols/pegp.pdf
http://jxb.oxfordjournals.org/content/51/350/1555.full.pdf+html
http://bmcgenomics.biomedcentral.com/articles/10.1186/1471-2164-14-216
There is a clear protocol that i suggest, Peg solutionis added at room temperature and the excess poured off after equilibratin, as in https://www.google.it/url?sa=t&rct=j&q=&esrc=s&source=web&cd=1&ved=0ahUKEwiDjraXzsDPAhXKAcAKHSK6AyUQFggeMAA&url=http%3A%2F%2Ffaculty.ucr.edu%2F~jkzhu%2Farticles%2F2006%2FProtocol%2520I-%2520Preparation%2520of%2520PEG-infused%2520plates.doc&usg=AFQjCNEaubpD74vS6__9faf9_BSQBIWqyw&cad=rja
I've done the above method that you explain in which you pour the PEG over the solidified media and that seems to be the best way to actually do that. The protocol provided by Guido seems like the way I have done it.
Hi! do you know who is the author of that protocole? I want to cite it in my thesis but I dont know how without that information!
thank you
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