Summary: 1
Ribonuclease protection assay
Aim : Analyze RNA 3'enc.
Solutions :
Hybridization buffer 5X : 100 ml Solution stock
200 mM PIPES pH 6.4 20 ml 1 M
2 M NaCl 40 ml 5 M
2 mM EDTA pH 7.5 2 ml 250 mM
DEPC H2O 38 ml
Ribonuclease digestion buffer : 100 ml Solution stock
10 mM Tris-HCl pH 7.5 1 ml 1 M
300 mM NaCl 6 ml 5 M
5 mM EDTA pH 7.5 2 ml 250 mM
DEPC H2O 91 ml
RNA loading buffer : 10 ml Solution stock
80% (v/v) formamide 8 ml 100%
1 mM EDTA 40 µl 250 mM
0.1% BromoPhenol Blue 1 ml 1%
0.1% XCB 200 µl 5%
DEPC H2O 760 µl

Source: Abou Elela, Sherif - Département de Microbiologie et d'Infectiologie, Université de Sherbrooke

Collections: Biology and Medicine