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northern blotting low copy mRNA [View Printable]
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Aaron Amano
Group: Member
Posts: 3
Joined: Aug 24, 2005
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I am planning on doing northerns for very low copy message. Would it be possible to do the northern using total RNA or would I have to purify mRNA? If I have to purify, any recommendations on kits out now?
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| Posted Oct 06, 2005, 17:23 PM |
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Sandy
Group: Member
Posts: 117
Joined: Nov 23, 2004
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| Aaron Amano said: | | I am planning on doing northerns for very low copy message. Would it be possible to do the northern using total RNA or would I have to purify mRNA? If I have to purify, any recommendations on kits out now? |
Your answer is yes, it is possible to run a northern with total RNA even for low copy number. Load at least 10ug of total RNA per lane, I have gone up to 20ug. however, it is also possible to isolate the mRNA. A good kit is the Qiagen Oligogex mRNA kit cat#70022. In my experience, I have gotten better northern/hybridization results using total RNA. |
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| Posted Oct 07, 2005, 15:09 PM |
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saswati1
Group: Moderators
Posts: 170
Joined: Mar 22, 2006
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Yes, it is possible | Aaron Amano said: | | I am planning on doing northerns for very low copy message. Would it be possible to do the northern using total RNA or would I have to purify mRNA? If I have to purify, any recommendations on kits out now? |
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| Posted May 31, 2006, 1:23 AM |
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