Thermo Fisher Scientific
FISH Tag™ DNA Multicolor Kit
F32951
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The goal was to detect 3D chromatin interaction between different chromosome. In order to distinguish various chromatin, I need to label them with different fluorescent dye. This kit helps to label BAC/Fosmid probe with different fluorescent dye.
DNA fluorescence in situ hybridization
HUVECs
The BAC/Fosmid probe were firstly digested with DNase I and nick translation was performed. After DNA precipitation overnight, amine-modified DNA was further labeled with fluorescent dye.
DNase I concentration and digesting time is the tricky part.
I closely followed the product protocol and found it's relatively easy. Since DNA FISH itself is a complicated experiment, the probe labeling step is crucial. I have use Green and Red dye performing DNA FISH on HUVECs and acquired excellent results.
N/A
Easy to follow and trouble shooting
Relatively expensive.
In general, this labeling kit is user-friendly, easy to follow and most importantly, able to trouble-shooting based on their suggestions.