Site directed mutagenesis

mutagenesis lecture - This lecture explains about the site directed mutagenesis including other techniques of mutagenesis like site specific recombination and transposition.
Site-directed mutagenesis is a molecular biology procedure that's used to make particular and intentional changes to the DNA sequence of a gene and any gene products.
Site-directed mutagenesis is likely one of the major techniques in laboratory for introducing mutation into a DNA sequence.
This video lecture deals with the mechanism and steps of site directed mutagenesis and the importance of side directed mutagenesis.
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[@thesahilstuff]

its 2023 still no-one's got this much clear and good explanation on this topic

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Glad it helped

[@flyingofdays]

I'm doing a reu using a lot of biochemistry and this is one thing that we will be doing. I have never taken a biochemistry course before so your videos have been a life saver in helping me understand some of the papers I have been reading.

[@HeterosexuaI]

I'm currently taking an upper-level microbiology research course and so much of the course material seems to fly over my head as though I have never heard about anything she is talking about. So far, your videos have made it much easier for me to follow along. Thanks, Shomu.

[@netaburnum1359]

Great lecture on the topic of site directed mutagenesis, thanks for the share, @Shomu's Biology.
Alternatively, it is used for investigating the structure and biological activity of DNA, RNA, and protein molecules, and for protein engineering. ---by Profacgen.

[@subhashakula1018]

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[@shomusbiologyofficial]

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[@visiblur]

Wherever my professor lacks, an Indian man will step up and teach me.

[@shomusbiologyofficial]

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[@aguardsjohnson8942]

Thank you so much, I like the way you articulate your explanation with thorough explanation.

[@harshabharwani4394]

Very well explained. Doubts clarified. Thank you!

[@tapelord2513]

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[@shomusbiologyofficial]

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[@hinasaleem9684]

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Sir i am a biotec student of class 12 and only your channel helps me out in clearing my biotec based douts. Thanks sir

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I am also biotech student

[@spulwasser]

Nice work ! Now that I understand what was meant with "nicks", I can also imagine how the thing works with deletions and insertions. Thx a lot

[@shomusbiologyofficial]

You're welcome

[@lanoanhduongthi3536]

Explain very easy to easy understand. Thank you sir

[@siroomar4711]

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[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@aldocastellani3536]

very helpful video. Very clear as usual. Thanks Shomu!

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You're welcome. Glad to hear that you're getting benefit from my lectures. Please subscribe and share

[@xxFabulous24xx]

You explain so clear, thank you so much !

[@elvisshiripindamhlanga123]

i like your teaching man its so splendid

[@shivpuranupay5840]

Beautiful explanation as always

[@TheUsermam]

Thank you for nice explain. I'm just start study about this topic and I'm so confuse. If you and your team have enough time can you explain about Iterative saturation mutagenesis?
Thank you

[@user-eg9xc8rr6c]

Just watched 1 hour before the paper and it helped me alott♥️👏

[@shomusbiologyofficial]

All the best

[@chriscarbe1042]

Very clear, Thank You!

[@hebaalghol4497]

Thank you very much.
I would like to ask if you do have a video about Triplet repeat primed PCR (TP-PCR), or if you can recommend a book or website that might help me understand it in details and thank you in advance.

[@marammokhtar1799]

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[@shomusbiologyofficial]

Glad to hear that you are getting benefit from my lectures

[@sharvanisivakumar4792]

Thank you SO much - this was so clearly explained!!

[@shomusbiologyofficial]

You're welcome

[@elijahmbiriyakura4153]

Thank you. good presentation

[@andresparra747]

Hmm, I think for this overlapping PCR you forgot to talk about kunkel or Quickchange approach in more detail, downstream treatments such as DpnI or enzyme digestion are quintessential for the application and sometimes is better to add the disadvantages such as mispriming or primer dimer hybridization. A part from that you are always in every search on youtube.

[@malabikabhowmik7933]

Very well explained, thank you so much Sir

[@shomusbiologyofficial]

You're welcome

[@raquel838]

muchas gracias aunque no hablo imgles me ayudo mucho....

[@bhuvanaanandhan548]

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[@shomusbiologyofficial]

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[@ivannabby15]

This is so helpful!! thank you for making this video

[@shomusbiologyofficial]

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[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

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[@annysentertainment1909]

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[@ammararaza3798]

Your way of reaching is amazing....really💞

[@shomusbiologyofficial]

Thank you

[@aratimaurya8464]

Nice explanation...I am mathematics student in Bsc. Now doing Ph.D in biochemistry so I have lots of problems in basic of biology...please suggest something how I clear my doubts?????😔😔😔

[@anythingsahaj7467]

i see a drastic transition your course subjects ? any particular reason for biology preference ?

[@keerthanak4171]

Ur explanation is simply awesome sir...😍😍

[@shomusbiologyofficial]

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[@ranishineraju]

I'm really a big fan of you sir..!!

[@shomusbiologyofficial]

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[@aayushpandey3122]

First video in I love it your study R and d then I subscribe and for follow in like you like on your videos good man

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@hamzamasood9933]

bht acha explain keya h AP NY , but kindly types of mutagenesis bhi add kijye , thank u

[@esmeraldasish]

Thank you man.

[@rafiullah8351]

AOA dear Shomu! What will be the result if only a single primer is used with a change in a single nucleotide please. Thanks

[@factfactnomi8682]

great job!

[@srikrishnanarasimhan9350]

Happy Teachers days SHOMU . love you .ummahhhhhh

[@shomusbiologyofficial]

Thank you

[@user-wi1fg5rc4m]

Good job bro!

[@healthmat3971]

You are the best sir. Thank you

[@shomusbiologyofficial]

You're welcome

[@sunitamohanty8499]

Thanks for such an amazing explanation sir ❤

[@shomusbiologyofficial]

You're welcome

[@nickrhoades2228]

Would the nicks cause a transformation efficiency decrease? Would it be better to use T4 ligase prior to transformation?

[@manognakuchibhotha7018]

sir i have a doubt that in the video at around 4:28 you have said that its different primer sequencing and is not same as PCR but again at last of the video you had presented it as a PCR method..... is it a PCR based primer designing method or no?

[@amangandhi1182]

Shomu Sir, please come to USA and start teaching the students in this manner. I am very sure that you are better than the teacher here

[@swastikmukherjee9648]

can I introduce 3-5 point mutations in this way? Also, do i have then design overlapping primers?

[@s.m.azadulalamraz8567]

great....go on

[@bharathraj5862]

Sir, what is the difference between the Site Directed Mutagenesis and Single Nucleotide Polymorphism?
Please clarify

[@a.k.m.zakirhossain6812]

It's actually a fantastic vedio 😊😊😊

[@shomusbiologyofficial]

Thank you

[@atmikapaul3049]

Do you also provide material for GATE prep sir?

[@kashafseducationalsite363]

one confusion sir.. how will this primer bind with dsDNA..? do we have to first denature the dsDNA of vector.. as usual occur in DNA replication...?

[@kunaalgoswami3581]

Thanks for the awesome explanation ✨

[@shomusbiologyofficial]

You're welcome

[@XY-yg1ci]

so helpful, make a difficult thing much easier

[@shomusbiologyofficial]

You're welcome

[@rishabhchandna787]

Sir can u explain how the primer will bind to the double stranded DNA ?????

[@manjusreemukherjee798]

Thank you so much...

[@shanishah4209]

Thank you sir for every thing

[@shomusbiologyofficial]

You're welcome

[@marjansiraj8030]

Congratulations for 2M subs sir!!!

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[@shomusbiologyofficial]

You're welcome. Glad to hear that you're getting benefit from my lectures

[@AbhishekPatel-ts6jp]

thanks a lot

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watched this for my upcoming biotech exam on july 20th !

[@srcreations8350]

So how u wrote ur exam?

[@shreyassehgal6849]

It was fine

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@@shreyassehgal6849 i think u wrote ur exam in online?? Isn't it ?

[@shreyassehgal6849]

Yes

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R u from I mean where r u from?which group u taken??

[@ezendianefojosiemkparu1951]

TX DEAR 4 HELPING MY CEREBRUM

[@yayahwinarti95]

I still wonder, if we would like to see the effect of mutation of a human gene, so does it mean we should insert the gene first on the vector and then we design a mutated primer for this gene?

[@julikavolkmann288]

could you please make a video about signature-tagged mutagenesis? :)

[@shashikalan7435]

Excellent teaching thank you

[@shomusbiologyofficial]

You're welcome

[@sahabuddinsahabuddin6216]

Thank you very much !

[@shomusbiologyofficial]

You're welcome

[@rasikakute537]

Very nice .. thank you!

[@shomusbiologyofficial]

You're welcome

[@akashvani3836]

Sir, please make a video on conventional mutagenesis

[@vibez9630]

is the klenow fragment that seals the nick instead of ligase?

[@debbiesoto4664]

tysm ly Shomu

[@shomusbiologyofficial]

You're welcome

[@m.hamzaramzan1283]

bro you are amazing.
Respect from Pakistan

[@shomusbiologyofficial]

Thank you. Glad to hear that you are getting benefit from the videos

[@nilayghosh5948]

English samajh mein ata hain pakistani o ko?😂

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@@m.hamzaramzan1283 I was thinking that about you.. PKMKB

[@m.hamzaramzan1283]

@@nilayghosh5948 hahahahahahaha that too was very creative😂😂. Why is there so much hate in your heart bro? Nationalism is the most toxic ideology and education is the only antidote for it. People in India get far better education than Pakistan but still there are a lot of people who cannot see past the borderline of imagination your rulling elite has set for you.

[@sunmathyk2360]

please do videos on PCR based site directed mutagenesis & cassette mutagenesis

[@kashikachowdhry2078]

Thankyou so much sir

[@shomusbiologyofficial]

You're welcome

[@Healthy_couple]

It's 2024 and it's still the best 😊

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@swatimishra7565]

Thanks sir.... It's really very helpful for me

[@shomusbiologyofficial]

You're welcome

[@asimkumarroy7471]

Sir, if you elaborate the full and another steps of Site Direcred mutagenesis. It will must help us to a broad idea about these topic. Thank you.

[@shomusbiologyofficial]

Okay

[@lakshmiamruthavalli7791]

Tnqu sir 👨‍🏫👨‍🏫

[@twisampatidas9965]

Excellent ❤🎉

[@shomusbiologyofficial]

Thank you

[@Abir_32133]

However , sir pls make a detailed video on pcr based site directed mutagenesis including.. overlap extension method,megaprimer PCR ,incerse PCR

[@shomusbiologyofficial]

Okay

[@kcampbell9248]

Hi Shomu how would you answer this question? "DNA sequencing suggests that random mutations in the spike protein of SARS-CoV-2 allow the virus to infect different hosts. Discuss a strategy you could use to test this theory"

[@shomusbiologyofficial]

To test the theory that random mutations in the spike protein of SARS-CoV-2 allow the virus to infect different hosts, one could use the following strategy:
1. **Sequence Analysis:**
- Collect viral samples from different hosts known to be infected by SARS-CoV-2.
- Perform high-throughput DNA sequencing of the spike protein genes from these samples.
2. **Mutation Identification:**
- Compare the sequences obtained from different hosts to identify any mutations present in the spike protein.
- Catalog and analyze the frequency and distribution of these mutations.
3. **In Vitro Mutagenesis and Protein Expression:**
- Use site-directed mutagenesis to introduce the identified mutations into the spike protein gene in a controlled manner.
- Express the mutated spike proteins in a suitable cell line.
4. **Binding Affinity Studies:**
- Perform binding affinity assays to test the ability of the mutated spike proteins to bind to the ACE2 receptors (or other relevant receptors) of different species.
5. **Infection Models:**
- Develop or use existing in vitro (cell culture) and in vivo (animal) models to test the infectivity of pseudoviruses or viral vectors expressing the mutated spike proteins.
- Assess the capacity of these viruses to enter cells from different species.
6. **Comparative Analysis:**
- Compare the infectivity data of viruses with and without the spike protein mutations across different host models.
7. **Statistical Validation:**
- Use statistical methods to determine whether there is a significant correlation between the presence of certain mutations and the ability of the virus to infect diverse hosts.
8. **Evolutionary Study:**
- Conduct a phylogenetic analysis to understand the evolutionary trajectory of the mutations and predict potential host jumps.
9. **Ethical and Biosafety Considerations:**
- Ensure all experiments are conducted within the ethical guidelines for the use of animals and biosafety levels required for SARS-CoV-2 research.
This strategy combines molecular biology techniques, comparative genomics, and functional assays to test whether mutations in the spike protein correlate with host range expansion in SARS-CoV-2.

[@suyashsachan74]

Loved it

[@shomusbiologyofficial]

Thank you

[@dipikasonkar8565]

sir if we will change the particular sequence for mutagenesis their is a possibility of occurence of some genetic disease also ?

[@shraddha8098]

Thank you ❤️

[@shomusbiologyofficial]

You're welcome

[@archanasatheesan7064]

Sr plzz upload a class on clb method of mutation detection

[@kashafseducationalsite363]

well explained .. from Pakistan..

[@shomusbiologyofficial]

Thank you so much for appreciating my efforts

[@writerexpertqueen1724]

Thank you sir👍

[@shomusbiologyofficial]

You're welcome

[@surjitdeka7092]

Other parts are available or not sir

[@elvisshiripindamhlanga123]

i am doing great in recombinant Dna technology now

[@selenophilicguy7635]

Poode myra

[@jsingh265]

Thanks bro.

[@shomusbiologyofficial]

You're welcome

[@Abir_32133]

Sir how two newly synthesized strand are combined🤔..

[@sidrakhan8589]

How to separate the mutated double stranded Dna from the parental dna that of plasmid???

[@Vikram798]

Awesome 😎😎

[@shomusbiologyofficial]

Thank you

[@user-go6xz4rb3f]

Thanksss for video

[@shomusbiologyofficial]

You're welcome

[@carlostettey2889]

How do i set the thermocycler to achieve this mutation?