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choice of in situ probes - (Jun/02/2005 )

Hi,
I hope someone can help me. I'm planning to do a lot of radioactive in situ's in the near future. But I don't know how to decide whether to use oligo's or cRNA probes?

I've also heard of this company, called GeneDetect, were you can have your oligo's designed and synthesized. As anybody tried this? And are there also other companies that design oligo's?

Thx a lot!!!

-Little LabGirl-

QUOTE (Little LabGirl @ Jun 2 2005, 04:27 AM)
Hi,
I hope someone can help me. I'm planning to do a lot of radioactive in situ's in the near future. But I don't know how to decide whether to use oligo's or cRNA probes?

I've also heard of this company, called GeneDetect, were you can have your oligo's designed and synthesized. As anybody tried this? And are there also other companies that design oligo's?

Thx a lot!!!



Hi

Yes we used the GeneDetect custom probe design and synthesis service a few years ago and had good results with our radioactive in situ hybridization. You can also follow the method in our manuscript if you like (see below) but it is essentially the same as the free protocol on their website

http://www.genedetect.com/protocols.htm

http://www.ncbi.nlm.nih.gov/entrez/query.f...2322&query_hl=7

Bye

-Scientist2002133-

QUOTE (Little LabGirl @ Jun 2 2005, 01:27 PM)
Hi,
I hope someone can help me. I'm planning to do a lot of radioactive in situ's in the near future. But I don't know how to decide whether to use oligo's or cRNA probes?

I've also heard of this company, called GeneDetect, were you can have your oligo's designed and synthesized. As anybody tried this? And are there also other companies that design oligo's?

Thx a lot!!!


is there any chance of doing Fluorescence ISH at your lab?? it's more sensitive and allows multiple target detection .... And if people are doing it in the lab all the equipment is there.
we by the way use oligoes synthesized by Eurogentec (europe)

-FISHer-

Hi again

I think most people would argue that radioactive in situ hybirdization is more sensitive than FISH (fluorescent ISH). At least this is my personal observation.

Thats not to say you can't get good results with FISH though of course and getting dual color FISH to work to localize 2 mRNAs at the same time would be great of course.

We liked using GeneDetect because of the probe design service they offer and as you can see from our publication it worked good for us for radioactive ISH.

Most oligo supply companies don't specialize in designing probes for ISH.

Good luck.

Bye.

-Scientist2002133-

Thx for your reply!

I have been doing double fluorescent ISH in the lab were I previously worked, and it's great. We do have the necessary equipment but I'm going for genes that are very weakly expressed so therefore I think I'll be using radioactive in situ's.

I think I'll try the GeneDetect design.
Thx again!

QUOTE (Scientist2002133 @ Jun 9 2005, 08:23 AM)
Hi again

I think most people would argue that radioactive in situ hybirdization is more sensitive than FISH (fluorescent ISH). At least this is my personal observation.

Thats not to say you can't get good results with FISH though of course and getting dual color FISH to work to localize 2 mRNAs at the same time would be great of course.

We liked using GeneDetect because of the probe design service they offer and as you can see from our publication it worked good for us for radioactive ISH.

Most oligo supply companies don't specialize in designing probes for ISH.

Good luck.

Bye.

-Little LabGirl-