Northern Blot

( www.abnova.com ) - The northern blot is used to study gene expression by detection of RNA (or isolated mRNA). A general blotting procedure starts with the separation of RNA by electrophoresis. The RNA can then be transferred to a nylon membrane through a capillary system and detected with a hybridization probe. More videos at Abnova www.abnova.com

[@youraffirmations2458]

The video is 13 years old, protocol has updated since then. Those days this was the method. Today the protocol is modified so follow the latest procedures. Overall, the basic principle will always remain same. ✌

[@brassicapanda6342]

Why put the RNA-side-membrane down in the UV crosslinking step? Does it matter? Shall I dry the membrane before the UV crosslinking?

[@권준-e4w]

RNA must be fixed by UV cross-linking because baking does not enhance signal intensity on Immobilon-Ny.

[@prisca2954]

I also have this question from watching the video. I checked the Stratalinker manual, and it suggests that the nucleic acid side should face up (opposite of video) and the membrane should be damp but not wet. See details here, pg 6: ipmb.sinica.edu.tw/microarray/index.files/Stratalinker%20UV%20Crosslinker.pdf Edit: The direction will depend on your UV light source. If using a transilluminator where the light comes from the bottom, than the RNA side should face down. If using a light source coming from above, than the RNA side should face up. Source: www.thermofisher.com/us/en/home/references/ambion-tech-support/northern-analysis/tech-notes/membrane-transfer-and-crosslinking-for-rna.html

[@youraffirmations2458]

@@prisca2954 The video is 13 years old, protocol has updated since then. Those days this was the method. Today the protocol is modified so follow the latest procedures. Overall, the basic principle will always remain same.

[@youraffirmations2458]

The video is 13 years old, protocol has updated since then. Those days this was the method. Today the protocol is modified so follow the latest procedures. Overall, the basic principle will always remain same.

[@vexanalasea]

Hi. I have a question regarding the Northern and Southern Blot. I have watched both videos of ABNOVA1, and I can't really tell a difference. Are the only differences in both techniques the solutions and the material(DNA/RNA)? I myself am a 3rd year college student, studying to become a technician in biochemsitry at Zuyd University in Heerlen, the Netherlands, and for my interim I'm supposed to do PAGE, western, eastern and southern blots. Regards, Vexan

[@veyselkoca]

I guess that in southern blot we are cutting DNA fragments via restrictive endonuclease enzyme like (EcoR1, BamH1).But in Northern blot that we don't cut fragments.This is a difference between Norhern and Southern.

[@drsharmabotany6632]

Southern is for DNA and northern is for RNA

[@youraffirmations2458]

Yes , process of transfer is same. But target molecule, extraction of DNA and RNA is different. DNA extraction protocol is different than RNA. Even the agrose gel percentage can differ while gel electrophoresis. Principle remains same for all the blotting techniques.

[@bookofpeacebeonibrahim9806]

Hello .......Am I allowed to use your four videos at a university conference?.....4 blotting.... please 🙏🙏🙏🙏💐💐💐💐

[@youraffirmations2458]

You can use any videos done by others available on public domain, but you cannot say that you are performing in that video. You can use it only for refernce and give crefit to the video owner/channel.

[@bookofpeacebeonibrahim9806]

@@youraffirmations2458 thank you very much

[@julietabaglietto4418]

SHOW THE RESULTS!!!! PLEASE AND THANK YOU, LOVE YOUR VIDS BIG FAN

[@julietabaglietto4418]

FIRST SUSCRIPTION EVER!!! GO SCIENCE

[@simply-angie]

I think Northern for DNA and Southern for RNA with the same method

[@lincolnoliveira3041]

Matsui