An improved, rapid Northern protocol.
We report a simple, sensitive and rapid method for performing Northern blotting analysis which avoids the use of toxic chemicals such as formamide and glyoxal. This technique allows sensitive detection of various transcripts from total RNA samples varying from 10 microg to 2.5 microg. These samples were probed for GAPDH and VCAM mRNA message using this technique and show similar results as conventional Northern blotting methods. In addition, this protocol can be accomplished much faster than the traditional formamide/formaldehyde or glyoxal protocols. This protocol can be easily implemented by most laboratories using inexpensive and reagents less toxic than those commonly used for RNA analysis.
Kevil
C G
CG
Departments of Molecular and Cellular Physiology, Louisiana State University Medical Center Shreveport, Louisiana 71130, USA.
Walsh
L
L
Laroux
F S
FS
Kalogeris
T
T
Grisham
M B
MB
Alexander
J S
JS
D000818:Animals; D015152:Blotting, Northern / Q000379:methods*; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis*; D019010:Vascular Cell Adhesion Molecule-1 / Q000032:analysis
D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1