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文献和实验Using Cell‐ID 1.4 with R for Microscope‐Based Cytometry
of yeast and mammalian cells processed by Cell‐ID. (A ) From left to right: yeast in focus, slightly defocused (note the dark ring on the border of the cells), and the same cells after Cell‐ID has identified each one and traced their borders correctly
Single Primer ("Semi-Random") P
a biased ratio of Po and Pn. Make 3 reactions with varying [Mg++]: 15 µL H2O 3 µL 10x H, M or L-PCRB 3 µL 4 dNTPs @ 2 mM each 3 µL Po @ 5 µM 3 µL template diluted to about 1 ng/µL 3 µL "T/TS" @ 0.4 u Taq/µL -------- 30 µL
. 8. (use combined filtrate) filter through sterile cheesecloth at +4 oC 9. (use supernatant) add 50 mM KCl, 2 mM MgCl2, 0.5 mM ATP for polymerization; incubate for 30 min; stir for 30 min; adjust to 0.8 M KCl; centrifuge at 100,000 x g for 2 h
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