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===Examination of chromosome prep=== *After disrupt the cells with syringe (i did it 20 times), the solution was filtered with 5 a micron filter. *I then spin the solution at 5000g for 5 minutes. A small white pallet is visible at the bottom of the tube. I transfer 800ul of the supernatant to another tube (Sup1). *I resuspended the pallet with the remaining ~200ul supernatant, repeat the centrifugation, transfer all supernatant to another tube (Sup2), and resuspended the pallet with 100ul of CIB. *I stained the chromosome suspension as well as Sup1/Sup2 with SYBG I, and checked the morphology with IX81. **The chromosome suspension contains small highly compact chromosomes, plus some larger objects, which could be nucleus or aggregates of chromosomes. There are ~35 chromosomes in each image taken with 40x objective, which is equivalent to 21 millions chromosomes per mL. Since I made an 1:2 dilution for staining/imaging, the actual density is ~42M/ml. **Interestingly, Sup1 is not blank. There are as many less compact chromosomes (~35 per image). So the density is also ~21M/mL. Chromosome suspension Sup1 [[Image:2010-08-19-chr-prep-40x-2.png|350px]][[Image:2010-08-19-Sup1-40x-1.png|350px]]
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