I want to extract DNA from blood found in whatmann number 3 filter paper using qiagen kit. The kit lacks the buffer that lysates the blood prior to extraction. Is there a lysis buffer I could use to lysate the blood without affecting the quality of DNA such that I obtain sharp bands when I do PCR? Also these filter papers are several years old.
Cut the area of blood spot present on filter paper.Keep the blood spot containing filter paper in a buffer made up of Ammonium chloride, Potassium bicarbonate and EDTA. Then incubate at room temperature for 3-5 min. Remove the paper and use blood sample for isolation of DNA as per the standard procedure
If you please, specify to me the volume and concentrartion of each reagent to make the buffer.
Thank you
Dear Rihab,
U may Use Buffer ATL (Qiagen cat# 19076).
Please read the protocol below for good and excellent yields
Best Regards
Hi,
Thank you. I read the manual and found it very beneficial but my problem is that I don't have buffer ATL.what I need is a lysing buffer that would be suitable for use with the extraction kit.
Rihab, qiagen buffer compositions are proprietary, so I am afraid anyone can help you with this. There are several blood lysis buffer recipes, but nobody can assure you that they will work with the qiagen columns and then you will have the risk to loose your precious archival samples! Qiagen sells the buffers separately and they are not quite expensive. If you cannot afford to buy them, I would advise you to go for a different, completely manual protocol from the beggining to the end.
Hi,
Thank you for your valuable.It is most helpful and definitely I will try the methods in the paper and see where they get before considering ordering the ALt from qiagen.
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