In Situ Hybridization

This lecture explains about the in situ hybridization process and the role of in situ hybridization techniques in identifying specific DNA sequence.
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I have a question:
If I have wild type and mutant models and I wanna examine the expression of specific markers Should I prepare probes for each markers for both mutant and wild type ?

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Thank you Shomu! I would like to know how the 2 strands of the DNA is separated in this process? by heating the cell ?

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What exactly did you mean by freezing the mRNAs?And how can you separate the 2 strands of a dsDNA?

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[@RahulKumar-mm5vm]

I have a question regarding dna sequence using FISH. "Is it possible to identify a particular gene sequence (for example - Amel Y) using FISH if there is lot of depurination (loss of adenine , guanine) in interested gene sequence ?"

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Thanks! nice & simple way to learn. How much work area is required for FISH setup?

[@markj.7687]

FISH set up is basic and does not require much bench top space. a simple table top would work just fine.

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called hybridization?

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Sir, is this same as southern and northern blotting techniques?

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how do you know the complementary sequence of the gene i.e. the probe or bait?

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[@nithyaa5841]

Sir what about in situ hybridization using non radiolabeled probes??

[@abhasingh6959]

how do u arrest cell in metaphase...when done on slide? reagent and concentration please...Nd please elaborate on signals .

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We arrest them using freeze fracturing technique

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[@nehanasir3492]

How is the complementary strand to the target sequence removed before hybridization?
Because it may happen that during hybridization or cooling, the target strand again bind to the complementary sequence instead of probe

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Hello, can you explain about detecting abnormal chromosomes such as 18,21, by FISH?

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[@moonsevinci88]

Thank you. Plz how we can separate the strands ? in vivant