Pulsed Field Gel Electrophoresis (pfge)

This lecture explains about the pulsed field gel electrophoresis process also known as the PFGE that helps in the separation of large DNA fragments with better resolution by shifting the electric field in time intervals. Pulsed field gel electrophoresis is a procedure used for the separation of massive deoxyribonucleic acid (DNA) molecules through applying to a gel matrix an electric subject that periodically changes course. Ordinary gel electrophoresis tactics for separation of DNA molecules provided huge advantages for molecular biology research. However, it was unable to separate very gigantic molecules of DNA simply. DNA molecules larger than 15-20 kb migrating by way of a gel will virtually move together in a size-impartial method. At Columbia school in 1984, David C. Schwartz and Charles Cantor developed a variant on the usual protocol via introducing an alternating voltage gradient to fortify the resolution of larger molecules. This manner became often called pulsed-subject gel electrophoresis (PFGE). The development of PFGE extended the variety of resolution for DNA fragments by means of as so much as two orders of magnitude. The process for this process is fairly much like performing a normal gel electrophoresis besides that instead of constantly jogging the voltage in a single direction, the voltage is periodically switched among three guidelines; person who runs by way of the crucial axis of the gel and two that run at an perspective of 60 levels either part. The pulse times are equal for each and every direction leading to a net ahead migration of the DNA. For enormously colossal molecules (up to round 2 Mb), switching-interval ramps can be used that raises the heart beat time for every course over the direction of a number of hours-take, for instance, increasing the heartbeat linearly from 10 seconds at zero hours to 60 seconds at 18 hours.
This approach takes longer than common gel electrophoresis as a result of the dimensions of the fragments being resolved and the fact that the DNA does no longer move in a straight line through the gel. Even as more often than not small fragments can in finding their manner by way of the gel matrix extra with ease than massive DNA fragments, a threshold length exists above 30-50 kb where all significant fragments will run at the identical price, and show up in a gel as a single giant diffuse band.
Nonetheless, with periodic altering of subject course, the more than a few lengths of DNA react to the exchange at differing rates. That is, higher pieces of DNA will probably be slower to realign their charge when field path is transformed, even as smaller pieces will probably be faster. Over the path of time with the constant changing of guidelines, every band will to separate increasingly even at very big lengths. Thus separation of very large DNA portions utilizing PFGE is made viable.
For more information, log on to-
www.shomusbiolo...
Get Shomu's Biology DVD set here-
www.shomusbiolo...
Download the study materials here-
shomusbiology.c...
Remember Shomu’s Biology is created to spread the knowledge of life science and biology by sharing all this free biology lectures video and animation presented by Suman Bhattacharjee in KZbin. All these tutorials are brought to you for free. Please subscribe to our channel so that we can grow together. You can check for any of the following services from Shomu’s Biology-
Buy Shomu’s Biology lecture DVD set- www.shomusbiology.com/dvd-store
Shomu’s Biology assignment services - www.shomusbiology.com/assignment -help
Join Online coaching for CSIR NET exam - www.shomusbiology.com/net-coaching
We are social. Find us on different sites here-
Our Website - www.shomusbiology.com
Facebook page- / shomusbiology
Twitter - / shomusbiology
SlideShare- www.slideshare.net/shomusbiology
Google plus- plus.google.co...
LinkedIn - / suman-bhattacharjee-2a...
KZbin- / thefunsuman
Thank you for watching

[@Sunflower1405]

You should be named "Molecular Genius" aka "Molecular Genetics". I wish some of my PhD lecturers at my MSc course would watch your videos and learn how to simply and effectively teach this Unit.

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@Sunflower1405]

@@shomusbiologyofficial and so nice and thoughtful as you reply to every single comment. what a gentleman.

[@swetayadav28_13]

7 years ago video yet the clarity !!...👏🔥💯

[@shomusbiologyofficial]

All the best

[@kewinmalla6233]

shomu biology is an amazingly reliable tutorial site,i can find all my syllabus here. Thanks from NEPAL.

[@shomusbiologyofficial]

+Kewin Malla great. I am so glad to hear that

[@samuelforward8453]

Thanks from a Food Science student in the UK!

[@user-pr3wl6qi3w]

sir your videos are really helpfull to understand techniqes and method👍

[@shomusbiologyofficial]

You're welcome

[@kashafseducationalsite363]

i dont knw why whenever i need to clear my concepts i prefer u.. u r amazing sir ..

[@shomusbiologyofficial]

Glad you liked my lectures

[@akhilajayamon3432]

All ur videos are really helpfull for my studies, and thank u for that. U are blessed with a good teaching skill. A good way of communication and convenient explanation. The main thing its very easy to follow u sir. Really hats off. Videos regarding microscopy, chromatography and electrophoresis are awesome. Once again appreciating ur effort ❤️

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@user-vj2et1ch3r]

It was very helpful! Thank you and your lecture style including your pronounce style is really catchy. It really helped my understanding.

[@shamsarefin5680]

Ton's of thanks

[@shomusbiologyofficial]

Thank you

[@kalyanlohar476]

Thank you Sir... Thank you 👍👍👌💞💞🙂🙂🤗

[@shomusbiologyofficial]

You're welcome

[@vaishnavikhavate5092]

It was very helpful & simply amazing.tq sm for your lecture.

[@swetaxavier6290]

Thanks for the video. U always rock

[@shomusbiologyofficial]

You're welcome

[@BaybeeDoll15]

thank you! very helpful when you explain it step by step :)

[@dipti7604]

Just loved it ....very helpful vedio . Thank you soooo much.

[@amnasarwar39]

Help alot. Kindly make video on microchip electrophoresis and concepts related with it.

[@fatemehakbari2189]

thank you . it was really very useful just like always .

[@abhilashasharma6255]

VERY WELL EXPLAINED

[@harryporterfield5701]

I recognize one of your images as coming from the CDC's PulseNet website. Where are the other images from? Please cite your references.

[@akvaryumdunyas5594]

Thanks, very helpful

[@satabdeepanda6810]

Thank uuu so much sir.😊

[@shomusbiologyofficial]

You're welcome

[@Greanestbean]

Thanks!

[@shomusbiologyofficial]

+Jon Roe you're welcome

[@kalyanlohar476]

thanks Sir.

[@anacarolinaprado1521]

Hello, I am standardizing PFGE for some kind of fungus and my races are getting blurry, with no definite bands. I wish I could have your contact and exchange ideas about what I could change in my protocol.

[@lidyahandayani4744]

Very nice video... 1 question : why do we need to use a gel (plug) to digest DNA using restriction enzyme? Why dont we just put a DNA suspension + restriction enzyme, and then put them in wells and run in pulsed field gel electrophoresis?

[@jyotikashyap5368]

Lidya Handayani maybe to avoid the step invovling isolation of the entire genomic dna of the bacteria,will save time.

[@heartforall5853]

Lidya Handayani I was wondering the same, but I think maybe because a normal DNA extraction will sheer/damadge the DNA a little bit and the agarose is maybe to protect the DNA and let it move easier in the agarose gel?

[@pushpasharma1813]

Because during the extraction process some of the DNA fragments might get degraded . So to avoid this , we use agarose plugs.

[@EducateWomenEmpowerWomen]

Sir, is agarose gel electrophoresis also known as submerged DNA electrophoresis? Or it is another technique?

[@niveditharajendran5113]

Pls do a video on isotachophoresis

[@brainynanaagyeman]

nice one

[@AmarSingh-ms1kg]

Audio is audible from one side while using earphone.

[@bhargavireddy5730]

Sir.. I have a doubt.. When we change the direction of the current there is a chance for the DNA fragments to get migrated into the adjacent lanes? Dnt they mess up?

[@shomusbiologyofficial]

They never travel through straight lines. And pulse field gel is kind of messy

[@bhargavireddy5730]

@@shomusbiologyofficial so.. Ecah time we will run only one sample sir?

[@MeenuBagariya]

Nice👌👌👌👌✍️✍️✍️✍️✍️🙏

[@shomusbiologyofficial]

Thank you

[@heartforall5853]

what about the other cell components? will these still be visible in the gel?

[@shomusbiologyofficial]

+Shinju Aishiteru no. For different types of cell components, we need to run different types of gel

[@uar9596]

Why there’s no audio?

[@samrahahsan2337]

there is no voice in video why its so

[@vivekabhyankar5029]

good1

[@abhishekrana5933]

board pe jyada smjh ata hai

[@shomusbiologyofficial]

Board video on offer coming soon