Hello,
Does anyone have suggestions for mRNA isolation kits? What kits have you used and how was the quality?
Any advice is appreciated.
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Experience with mRNA isolation kits?
Started by sarabell, Jan 04 2012 06:04 AM
mRNA isolation kit RT-PCR
3 replies to this topic
#2
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Veteran
Posted 04 January 2012 - 11:04 PM
For total RNA isolation we have used RNeasy and it works great. Also used Promega total RNA extraction kit with far less yield. Not sure about kits for mRNA only but I hope this helps.
#3
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an elder
Posted 05 January 2012 - 11:49 AM
i think this is the one i used (a long time ago, i was building a library) and it worked well:
clontech
you can also check out this page from life technologies:
mrna extraction
clontech
you can also check out this page from life technologies:
mrna extraction
Edited by mdfenko, 05 January 2012 - 11:50 AM.
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
#4
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member
Posted 10 January 2012 - 08:17 PM
Typically trizol works well. 70-100mg of tissue will yield a LOT of mRNA.
RNaqueous kits from Ambion work beautifully if you have difficult tissues (especially high fat content ones). I have gotten great results from 2mg pieces of brain and from adipose, which doesnt have much RNA. There is a micro kit (1-10mg) and the regular kit with columns that go to 100mg of tissue. Homogenize then spin down any debris that might clog the filter. Generally the RNA is very pure but (nanodrop readings at 2 for both ratios) but sometimes there will be contamination/carryover, which is fixed with an overnight lithium chloride precipitation. For microarray (or anything you need super clean RNA for) do a DNAase treatment then LiCl precip (both come with the micro kit, LiCl comes with the regular kit).
RNaqueous kits from Ambion work beautifully if you have difficult tissues (especially high fat content ones). I have gotten great results from 2mg pieces of brain and from adipose, which doesnt have much RNA. There is a micro kit (1-10mg) and the regular kit with columns that go to 100mg of tissue. Homogenize then spin down any debris that might clog the filter. Generally the RNA is very pure but (nanodrop readings at 2 for both ratios) but sometimes there will be contamination/carryover, which is fixed with an overnight lithium chloride precipitation. For microarray (or anything you need super clean RNA for) do a DNAase treatment then LiCl precip (both come with the micro kit, LiCl comes with the regular kit).
