Hi there!
currently i am working with spin column kit for DNA extraction I've put almost 600 uL of buffy coat. after a lot of washing, the RBC's remained stucking in filter? How could i get rid from this contamination after extraction ?
There are a few things you can do:
-get better columns that cope with blood/buffy coats
-prior to adding the buffy coats to the columns, when you put the cell lysis buffer in with the buffy coats/blood sample, ensure you vortex the tubes thoroughly to enable sufficient breakdown of the RBCs and even distribution of the lysis buffer
-take more blood liquid/less buffy but at the expense of yield.
-add 96% ethanol after adding in the cell lysis buffer and vortex to get a less viscous mixture (risk lower yield but higher purity)
what is your purity like on the spectrophotometer?
Katerina Christofides thanks katerina for your advice I will try it. Do u know how i could solve my problem in the purity after extraction? such as these results
your concentration is good, try loading less buffy coat to the column.
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