RNA pellet - (Jan/07/2008 )

in RNA extraction ..is there any tricks or tips to visulize the pellet ... i am dealing with very small amount of RNA so am having hard time seeing the pellet and i think am loosing it while washing..

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When using isopropanol for RNA extraction you get quite a clear pellet. What I do is put a mark on the outside of the tube to highlight where the pellet should be before centrifugation And be careful not to disturb that area when you aspirate off your liquid.

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I use 1-2 ul of Glycogen for Precipitation. I normaly isolate RNA with Trizol. Before I transfer the upper (clear) phase into a new tube, I put 20mg of Glycogen into the tube. Afterwarts I add the upper phase, then the Isopropanol and then I vortex for aprox. 5 sec. and incubate for 5 min at RT.

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i checked for the glycogen ...and in one place they said is not recommended when dealing with sample rich with proteoglycan !! do you have idea about that? in matter of fact am dealing with ligament tissue, hence is rich with PG

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you could try something like pellet paint.

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try "glycoblue" to identify the tiny rna pellet as blue

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Yes the blue glycogen really helps visualize the otherwise semi-transparent pellet when dried. Another carrier you can use is linear acrylamide.

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