CSCL gradient centrifugation.

: Read some information about CSCL gradient centrifugation, but feel different, and the process is not clear. Now I would like to ask some questions:
1, CSCL density and number of turns.
2, about CSCL sterilization.
: Because it is to do in vivo experiments, thank you all this is our laboratory operation steps:
in every 4 ml dissolved gradulate TE solution, add 4 g of CSCl, hand temperature
heating
so that CsCl solid solution solution, and then transfer to the overspeed
centrifuge tube
, add 0.2 ml of EB (10 mg/ml) to each tube, and then adjust the balance with TE, so that the liquid surface distance from the tube mouth is about 1 mm.
25 degrees C, 62 500 rpm, centrifuged 16 hrs. After centrifugation is complete, gently remove the centrifuge tube, fixed to the iron frame, under the dark room, with a 5 ml disposable syringe with 18 needles in the nearest red ring belt (ultra-helix granules belt) near the pipe wall, the purpose belt slowly sucked out, over-the-top try not to layer the liquid surface disturbance.
transfer the liquid containing the ultra-helix granules into the new overspeed centrifuge tube, add a TE solution containing 1 g/ml CsCl, add 0.05 ml of EB (10 mg/ml), 25 degrees C, 625,000 rpm, centrifuge 16 hrs.
.