Hi Colleagues,
Molecular Combing enables the analysis of single DNA molecules on a glass slide. The DNA molecules are fixed on the glass slide and oriented like a line or fibre on the slide. Following this one can apply fluorescence probes to visualize chromosomal abberations like in FISH technique.
My question is know, do have somebody experience with this technology?
What is your application and why you are not doing NGS or FISH?
Thank you all in advance for your help!
Hi Martin,
When I was a graduate student my lab had joint lab meetings with another that was attempting to set up this technique, so they talked about it extensively. They were interested in looking at origin of replication firing, so they actually performed BrdU incorporation and sometimes FISH alongside the combing. I think for their purpose, molecular combing was ideal because they could actually monitor number and location of origins of replication. I am sure there are other applications, but this is the only one that I am familiar with. For this purpose, you can see why NGS or FISH alone wouldn't work. Hope this helps.
Hi Andrew!
Thank you for your reply.
Yes, I think so too, that NGS is not able to solve this.
Do you think molecular combing could be a diagnostic tool for cancer? I am asking because of analysis of single circulating tumor cells would be interesting for me.
From what I know about the technique, it is fairly challenging and only a few labs use it on a regular basis. Also, I believe that you need large amounts of DNA, so it wouldn't be suitable for a single cell analysis. If you are looking for a diagnostic tool for cancer, your best bet would definitely be identifying a marker for that particular cancer line. I don't know if examining genome dynamics (ie., with FISH or combing) would be sensitive/fast enough to serve as a diagnostic tool.
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