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Qiagen mini prep cell fragment pellet


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#1 fred1234

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Posted 03 July 2013 - 08:49 AM

Hi,

I fund when using the Qiagen miniprep kit, after lysis and then neutralisation with N3, that the cell debris I get is impossible to pellet. It forms a weird semi-solid that floats between what seems to be two different phases. It really limits the amount of supernatant I can get.

I remember experiencing this before but it's particularly bad at the moment for me. Any advice much appreciated.

#2 bob1

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Posted 03 July 2013 - 12:23 PM

Ensure that you mix the lysis and neutralization buffers well, spin at greater than 10,000 RCF.

#3 phage434

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Posted 03 July 2013 - 03:19 PM

This can be caused by large pellets. You may want to reduce the volume of cells, or grow in LB instead of a rich medium.
You can always filter with a coarse pore filter, such as cheese cloth.




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