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Chromosome Replicating Timing Combined with Fluorescent In situ Hybridization

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17:14 min

December 10th, 2012

December 10th, 2012

13,846 Views

0:05

Title

1:53

BrdU Incorporation: Terminal Labeling

2:35

Chromosome Harvest of Monolayer Cells Cultures

5:53

RNase Treatment and Ethanol Dehydration

6:38

Fluorescent Labeling BAC DNA: Nick Translation

8:19

In situ Hybridization

12:18

Capturing Images and Quantifying BrdU Incorporation

14:27

Results: Chromosome Replication Timing and FISH

16:45

Conclusion

A quantitative method for the analysis of chromosome replication timing is described. The method utilizes BrdU incorporation in combination with fluorescent in situ hybridization (FISH) to assess replication timing of mammalian chromosomes. This technique allows for the direct comparison of rearranged and un-rearranged chromosomes within the same cell.

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