DENATURING AGAROSE GEL FOR RNA
Posted 14 January 2010 - 09:20 PM
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,
Posted 17 January 2010 - 03:17 PM
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 editions are good, though some a little outdated.
Edited by bob1, 17 January 2010 - 03:18 PM.