A rapid and simple one step method for isolation of poly(A)+ RNA from cells in monolayer.
A simple method was developed to isolate low abundance hormone receptor poly(A)+ RNA from cells in tissue culture. Adherent cells in tissue culture plates were directly released with proteinase K and solubilized in SDS. Oligo(dT) cellulose was directly added to the lysate to obtain poly(A)+ RNA. Yields and purity of the poly(A)+ RNA were comparable to other more lengthy methods. IGF-I receptor and insulin receptor mRNA could be detected on Northern blot without any degradation.
Hartmann
K K
KK
Diabetes and Endocrine Research Department, Mount Zion Hospital and Medical Center, San Francisco, CA 94120.
Papa
V
V
Brown
E J
EJ
Doerries
U
U
Rosenthal
S M
SM
Goldfine
I D
ID
D000818:Animals; D001943:Breast Neoplasms / Q000032:analysis; D002460:Cell Line; D015342:DNA Probes; D005347:Fibroblasts / Q000032:analysis; D006801:Humans; D051379:Mice; D009132:Muscles / Q000032:analysis; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*; D012333:RNA, Messenger; D011972:Receptor, Insulin / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000235:genetics*; D017451:Receptors, Somatomedin; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D011061:Poly A; D012313:RNA; D011972:Receptor, Insulin