Simultaneous and rapid isolation of bacterial and eukaryotic DNA and RNA: a new approach for isolating DNA.
A very simple procedure for the simultaneous preparation of genomic DNA and total RNA is described. The procedure is essentially the same for eukaryotes and prokaryotes except for the lysis buffer and can be used for small or large numbers of cells. Mammalian cells are lysed in sodium dodecyl sulfate and bacterial cells are lysed in Triton X-100, both in the presence of EDTA. RNA is obtained in the aqueous phase after phenol (acidic pH):chloroform:isoamyl alcohol extraction. DNA is eluted out of the organic phase (and the interface) into the aqueous phase by increasing the pH with highly basic 1 M Tris solution. The method is extremely rapid for small or large numbers of cells, and several large samples can be processed in one day. The qualities of both nucleic acids are excellent and the yield is high.
Majumdar
D
D
Department of Biology, Brown University, Providence, RI 02912.
Avissar
Y J
YJ
Wyche
J H
JH
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D005821:Genetic Techniques; D008322:Mammals; D012313:RNA / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000302:isolation & purification
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012313:RNA; D004247:DNA