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1. To which reagent does the first step of the protocol "Add...
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Wojciech Francuzik Jun 3, 2015
1. To which reagent does the first step of the protocol "Add 100 μl of yeast cell lysis buffer to a 2 ml safe-lock tube. " refer to?
Yeast cell lysis buffer is listed in the materials and reagents list as "(Epicentre, catalog number: MPY80200)". This is actually the MasterPure K Read more
Yeast cell lysis buffer is listed in the materials and reagents list as "(Epicentre, catalog number: MPY80200)". This is actually the MasterPure K Read more
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Julia Oh Author Answered Jun 4, 2015
National Human Genome Research Institute, National Institutes of Health, MD
Because of the low biomass in the skin, we try to keep the volumes low. This amount is adequate to moisten the swab for sampling a 4cm2 area. If you are planning on sampling a larger area, it'd be worth experimenting with larger volumes.
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0 comments ShareWojciech Francuzik Answered Jun 3, 2015
Charité Universitätsmedizin Berlin
Thank you for the prompt reply.
Just to be sure: after step B3:
"Short spin tubes, then open and add 1 μl of ReadyLyse and 250 μl of Yeast Cell Lysis Solution."
we will have 350 μl of Yeast Cell Lysis Solution in our tube.
Why are we only starting with 100 μl for the extraction? 100 μl is rather small volume to moisten the Catch All swab. I would normally use 1000 μl of the SCF-1 solution (described in the methodology of the HMP for sampling skin).
Could you shortly explain the rationale here?
Just to be sure: after step B3:
"Short spin tubes, then open and add 1 μl of ReadyLyse and 250 μl of Yeast Cell Lysis Solution."
we will have 350 μl of Yeast Cell Lysis Solution in our tube.
Why are we only starting with 100 μl for the extraction? 100 μl is rather small volume to moisten the Catch All swab. I would normally use 1000 μl of the SCF-1 solution (described in the methodology of the HMP for sampling skin).
Could you shortly explain the rationale here?
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0 comments ShareJulia Oh Author Answered Jun 3, 2015
National Human Genome Research Institute, National Institutes of Health, MD
Yes, this is the yeast cell lysis solution from the MasterPure Kit.
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