DENATURING AGAROSE GEL FOR RNA - (Jan/14/2010 )

Dear All,

Presently am isolating RNA from animal tissue cultures using the trizol method.

My questions are :

1. In the denaturing gel, while loading the sample the sample mixture has both formaldehyde and formamide, y do v hav 2 use both ? Wont formaldehyde alone do the job ? And also formaldehyde is already present in the gel, so y again in the sample mixture ?

2. If the native state of RNA is lost, how come EtBr intercalates with the RNA ?

3. Why r v using MOPS buffer ? Why not TAE or TBE buffer ?

4. Why r v seeing only the 28s, 18s rRNA on the gel and not mRNA or tRNA ??? How can the quality of mRNA b assessed by seeing the quality of rRNA ?? What is the correlation ??

Also suggesst some good n simple book 2 look for all d mol bio techniques n principles......


Hope am not askin too much !!!

Thanks in advance,


josh !

These look a lot like homework...

Have a go at answering them yourself and tell us what you come up with.

A good book is Molecular Cloning: a laboratory manual, by Sambrook, Fritsch and Maniatis. Any of the 3(4?) editions are good, though some a little outdated.