When I prepared plasmid, after added solution 3, centrifuged the lysate at 10,000g, 10 min, before pour the supernatant into column, I still can see some floating debris in the supernatant, how I remove the floating debris with any filter? Do you have any filter for recommendation? thanks.
Please look at the following below links which may help you in your analysis:
http://wang.ucsd.edu/protocol/2.%20molecular%20cloning/2.3%20Amplification/Shipping_and_Receiving_Plasmids_on_Filter_Paper.pdf
https://openwetware.org/wiki/Pelling:Protocols/Extracting_DNA_from_Filter_Paper
https://static1.squarespace.com/static/51cb031de4b08c2dba6581d3/t/53ff6dcde4b051bac941ad85/1409248717435/Shipping+of+plasmids+on+filter+paper+and+their+recovery.pdf
Thanks
Please have a look on the link below.
https://www.fishlore.com/aquariumfishforum/threads/what-is-the-floating-debris-and-how-to-remove-them-from-fish-tank.384739/
Hi Aiguo,
- If I understand correctly, you are facing a problem in getting clear supernatant after adding solution 3 (neutralization buffer) during Plasmid isolation. This may happen for two reasons, [1] After adding solution 2 (lysis buffer) if you need a gentle mixing either by rotating the tube by hand or gentle mixing by pipette and then add solution 3 (neutralization buffer) and again mix it gently without vortexing. If you are vortexing it, you will get these small suspended particles (cell debris) in the supernatant. [2] check if your solution 3, which is actually neutralization buffer with high salt is not very old.
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