This kit has been designed for isolation of total RNA from animal tissues, Cell cultures and blood. Tissue RNA Kit allows simultaneous processing of multiple tissue samples in less than 30 min. The procedure completely removes contaminants and enzyme inhibitors making RNA isolation fast, convenient, and reliable.
Some tissues such as muscular tissues, heart and aorta tissues make RNA isolation more difficult because they contain contractile proteins, connective tissue and collagen. These tissues need an intensive lysis process by homogenizer.
Lysis solution contains phenolic component to inhibit RNase activity and to remove DNA and protein content of the samples. It should be handled with care under a FUME HOOD.
After addition of chloroform, centrifuge of the lysate makes a three phase soltution, the same as Trizol method. Do Not Touch Interface or Phenol Layers. Try to remove upper part of aquous solution and left aquous solution near interface layer to decrease DNA cotamination of the samples. This ensure an efficient column purfication without closure of the silica columns.
Silica column step removes most of contaminant and give a pure RNA .
Highly denaturing phenolic buffer inactivates RNases and remove DNA content.
Procedure efficiency is not very sensitive to correct amount of the start sample. Most of the column based RNA purification kits should be optimized for a correct amount of samples. Higher amounts of samples decrease RNA yield and increase contaminants.
Combination of a column based procedure with a phenolic base solution method.
Without column closure due to oversample.
No precipitation step.
No phenolic or protein contaminants.
Procedure will take 30 minutes.