ISOLATION OF RNA FROM PLANTS - METHODOLOGY QUESTION

Ed Davis edd at OSUUNX.UCC.OKSTATE.EDU
Sat Nov 12 16:08:04 EST 1994


I AM ABOUT TO PURIFY TOTAL RNA FROM GOSSYPIUM (COTTON) AND AM PLANNING TO FOLLOW A METHOD OUTLINED BRIEFLY IN NUCLEIC ACIDS RESEARCH BY MALIYAKAL E. JOHN OF AGRACETUS, INC.(VOL 20, NO9, page 2381) WHICH IS
PREFERABLE FOR PLANTS WHICH ARE HIGH IN POLYPHENOLICS. BRIEFLY, THE RNA WAS GROUND IN LIQUID NITROGEN, HOMOGENIZED IN THE PRESENCE OF A BUFFER CONTAINING, AMONG OTHER THINGS, PVP, FILTERED AND THEN COLLECTED FOLLOWING CENTRIFUGATION IN CESIUM CHLORIDE BY THE METHOD OF CHIRGWIN. IT WAS THEN FURTHER PURIFIED BY PHENOL:CHLOROFORM EXTRACTIONS AND PRECIPITATIONS IN THE PRESENCE OF AMMONIUM ACETATE AS DESCRIBED BY CROUSE AND AMORESE FOR DNA (1987, FOCUS, VOL9, 3-5).  I AM HAVING TROUBLE OBTAINING THIS ARTICLE. I

F ANYONE IS FAMILIAR WITH THIS METHOD I WOULD APPRECIATE YOUR HELP AND ADVICE.  
ANY OTHER CONSTRUCTIVE ADVICE WOULD BE MORE THAN WELCOME.  THE RNA WILL BE USED FOR SPECIFIC AMPLIFICATION BY PCR.



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