a*****s
发帖数: 838 | 1 Need to separate some virions with Sucrose ultracentrifugation (PI said we
don't need to do CsCl :( - that would have been easier to search for
literature if so). Anyway, I did some trial runs. My goal is to separate the
samples to several layers (if there is different shape of viral particles).
Then collect the layers and run on western blot to see and compare.
take I : 17300 rpm at 4C, x 90 min --> no separation
take II: 30000 rpm at 4C, x 90 min --> no separation
take III: 30000 rpm at 4C, x 3 h ---> still no separation
the sucrose pads i used are: 20% (upper), 35% (middle); 50% (bottom); let
sit at 4C overnight before loading samples and ran the program.
total volume 2 ml (each layer 0.5 ml -- including sample layer). | l*h
发帖数: 4124 | 2 If you calculate the centrifugal force (in g) and know the sedimentation
rate of your virion, you can have an educated guess about the conditions you
need.
the
).
【在 a*****s 的大作中提到】
: Need to separate some virions with Sucrose ultracentrifugation (PI said we
: don't need to do CsCl :( - that would have been easier to search for
: literature if so). Anyway, I did some trial runs. My goal is to separate the
: samples to several layers (if there is different shape of viral particles).
: Then collect the layers and run on western blot to see and compare.
: take I : 17300 rpm at 4C, x 90 min --> no separation
: take II: 30000 rpm at 4C, x 90 min --> no separation
: take III: 30000 rpm at 4C, x 3 h ---> still no separation
: the sucrose pads i used are: 20% (upper), 35% (middle); 50% (bottom); let
: sit at 4C overnight before loading samples and ran the program.
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