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In Situ Hybridization for BEGINNERS 

RealizingResearch
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In situ hybridization is a technique used by scientists to label DNA and RNA, but why do we need to label DNA/RNA?
Here I explain the purpose of In Situ Hybridizations, provide a brief description of the two major types (chromogen and fluorescence in situ hybridization (FISH)), give several examples of their use, and explain some drawbacks.
This is a part of my Slice of Science series where I "briefly" describe some interesting scientific techniques.
Items used in the video:
Music: • Child of Time (1 HOUR)...
Articles used as examples:
Cancer Cell DNA Translocation Example
Louzada S, Adega F, Chaves R. Defining the sister rat mammary tumor cell lines HH-16 cl.2/1 and HH-16.cl.4 as an in vitro cell model for Erbb2. PLoS One. 2012;7(1):e29923. doi: 10.1371/journal.pone.0029923. Epub 2012 Jan 10. PMID: 22253826; PMCID: PMC3254647.
Figure 2A
Dentate Gyrus/Low Level Chromogen RNA In situ
Caramello A, Galichet C, Rizzoti K, Lovell-Badge R. Dentate gyrus development requires a cortical hem-derived astrocytic scaffold. Elife. 2021 Jan 4;10:e63904. doi: 10.7554/eLife.63904. PMID: 33393905; PMCID: PMC7806271.
Figure supplement 5A
Enthesis Example/Double RNA FISH
Kult S, Olender T, Osterwalder M, Markman S, Leshkowitz D, Krief S, Blecher-Gonen R, Ben-Moshe S, Farack L, Keren-Shaul H, Salame TM, Capellini TD, Itzkovitz S, Amit I, Visel A, Zelzer E. Bi-fated tendon-to-bone attachment cells are regulated by shared enhancers and KLF transcription factors. Elife. 2021 Jan 15;10:e55361. doi: 10.7554/eLife.55361. PMID: 33448926; PMCID: PMC7810463.
Figure 1A, Figure 2C
These articles are distributed under the terms of the Creative Commons Attribution License (CC-BY-4.0), which permits unrestricted use and redistribution provided that the original author and source are credited.
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Опубликовано:

 

16 фев 2021

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Комментарии : 30   
@surayuthpintawong8332
@surayuthpintawong8332 Год назад
Thanks, the way you described this material is so good.
@biofilia_
@biofilia_ Год назад
Very clear explanation. Thanks!
@maryg5941
@maryg5941 2 года назад
Super cool video! Watched it as a resfresher for a job Interview. You did a great job! 😍
@aliiranban8871
@aliiranban8871 2 года назад
I watched a this for a college presentation and it's nearly completely what i needed to know about it! I eagerly look forward for your new content. Maybe you could cover more techniques in microscopy.
@tah4155
@tah4155 2 месяца назад
Wow! Amazing teacher! thank you for this video :))
@Vislumbre.
@Vislumbre. Год назад
Heyyy, that was a great explanation, thank you very much!!
@airicsnider7572
@airicsnider7572 2 года назад
Thanks, Comrade.
@solz006
@solz006 Год назад
thanks a lot for the video! really informative
@farahal-najjar6480
@farahal-najjar6480 Год назад
Thank you!
@SirPlantsaLot
@SirPlantsaLot 2 года назад
'gene for people who like hummus' loooool, you're great at explaining things, thank you!
@amruthatk6371
@amruthatk6371 3 месяца назад
THANK YOU 🤝
@codeblue3228
@codeblue3228 9 месяцев назад
Wonderful explanation, please make a video on whole mount insitu hybridization.
@nileshsharma0212
@nileshsharma0212 2 года назад
Please make more videos about techniques which you use for your research
@tonysarris668
@tonysarris668 Год назад
Thanks for the video. Could flow cytometry be used for the detection of these probes?
@yelkayap7813
@yelkayap7813 10 месяцев назад
thank you so much for this! i feel like i can pass my exams tomorrow ^^
@anajuliamacedo6239
@anajuliamacedo6239 Год назад
YOU SAVED ME
@anupshrestha431
@anupshrestha431 Год назад
thanks for the clear explaination. Bit off topic but can you share the source of the background music? I think it made the video much nicer when it started around 6:30.
@imethan-youtubetips1770
@imethan-youtubetips1770 3 года назад
7:25 this was very informative. Also good editing. Thumbs up from a fellow creator 😅
@realizingresearch4194
@realizingresearch4194 3 года назад
Thanks! I'm really glad you liked it.
@jonnyjaap
@jonnyjaap 3 года назад
Nice video and you have a lot of positive energy, cool! You could talk about qPCR, and it can be used for quantification, right ?
@realizingresearch4194
@realizingresearch4194 3 года назад
I am definitely gonna do a video on qPCR. I just wasn't sure when. I'll move it up. Thanks for the nice comment!
@anapalacios8452
@anapalacios8452 2 года назад
¿What should I use in order to quantify the expression of the genes?. Thanks!
@realizingresearch4194
@realizingresearch4194 2 года назад
There are a few options, I think the best and most widely used tool is qRT-PCR.
@samuellevac1919
@samuellevac1919 2 года назад
What do you mean by temperature sensitive? More specifically how does the cold affect it?
@realizingresearch4194
@realizingresearch4194 2 года назад
Great question! Thanks for asking! The primary effect temperature has on this type of reaction is changing the binding between the probe and your DNA/RNA. The DNA and probe will not bind at too high of temperatures and too low of temperatures, say below 40C/104F, you have nonspecific binding. This is similar to where I described your probe binding to other RNA/DNA. So you need to find the optimum temperature for your reaction based on your tissue and probe. Does that make sense?
@samuelfink8569
@samuelfink8569 3 года назад
I wish I exhibited the "Super Cool Gene" :(
@realizingresearch4194
@realizingresearch4194 3 года назад
I mean, you did watch the video....
@addieandlogansyaya
@addieandlogansyaya 3 года назад
I wonder if she has sibling competition issues
@realizingresearch4194
@realizingresearch4194 3 года назад
Only in video games and school grades and life. 😁
@vincent70
@vincent70 2 года назад
You're funny
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