Answer the question below C RNA isolation working with RNA W

Answer the question below: C) RNA isolation/ working with RNA: What special precautions must be taken when working with RNA? Why? How do you know if you\'ve successfully isolated good quality RNA? (http://www.ambion.com/techlib/basics/rnasecontrol/index.html)

Solution

When we are working with RNA RNAse contamination is one of the problem that is commonly encountered as these RNAses are ubiquitous and are found practically found everywhere. These molecules are secereted in to the body fluids like tears , sweat, mucous etc. to dfend against the microsorganisms that invade the body.

The common precautions to be taken while working with rna is to prevent the possible contamination with RNAses to avoid the degradation of the molecule.They are as follows:

1. wearing of gloves throughout the experiment to prevent the RNases from hands and changing of the gloves when they come in contact with the door knobs or any body surface like skin.

2.Chemical reagents that are used should be free of RNAses.

3.Usage of RNASe free tips and tubes.

4.Lab surfaces to be cleaned with RNAse decontamination solutions like AMbions zapOr with mild solution of bleach or with NaOH.

5. glass weare and metal wear are to be autoclaved at  450°F for a duration of 2 hours.

6. Buffers and solutions to be used should be treated with DEPC to eliminate RNAse contamination.

The proof of good quality RNA being extracted is the presence of 28S to 18S ribosomal RNA band in the ratio of 2:1 when the sample is run on denaturing agarose gel with ethidumbromide staining.