[MA100] BioZol reagent

MSDS – BioZol RNA

Features : High purity RNA isolation (1.6-1.8 A260/280)

Reagents required, but supplied
Chloroform, Isopropyl alchol, 75% Ethanol (in DEPC-treated water), DEPC-treated water

Precautions for Preventing RNase Contamination
RNase can be ontroduced accidentally into the RNA preparation at any point in the isolation procedure
through improper technique/ Because RNase activity is difficult to inhibit, it is essential to prevent its
introduction. The following guidelines should be observed when working with RNA.
1. Always were disposable gloves. Skin often contains bacteria and molds that can contaminate an
RNA preparation and be a source of RNases.
2. Use sterile, disposable plasticware and pipettes reserved for RNA work to prevent cross-contami-
nation with RNases from shared equipment.
Troubleshooting Guide
1. Expected yields of RNA per mg of tissue or 1 x 106 cultured cells
Liver and spleen, 6~10ug Kidney, 3~4ug Placenta, 1~4ug
Epithelial cells, 8~15ug Fibroblast, 5~7ug Skeletal muscles and brain, 1~1.5ug
2. Low yield
Incompleted homogenization or lysis samples
Final RNA pellet incompletely redissolved
3. A260/280 ratio < 1.65
RNA sample was diluted in water instead of TE prior to spectrophotometerric analysis.
Sample homogenized in too small a reagent volume.
The aqueous phase was contaminated with the phenol phase.
4. RNA degradation
Tissues were not immediately processed or frozen after removal from the animal.
Cells were dispersed by trypsin digestion.
Aqueous solution or tubes were not RNase-free.
Formaldehyde used for agarose-gel electrophoresis had a pH below 3.5
5. DNA contamination
Sample homogenized in too small a reagent volume.
Samples used for the isolation contained organic solvents strong buffers, or alkaline solution.