Gene Expression Analysis and DNA Microarray Assays

Рет қаралды 352,104

Күн бұрын

If we want to understand a biological organism, we turn to the expression of its genome. Which genes are being expressed, and in which cells, and when? How does this differ between a normal cell and a cancer cell? We have incredibly sophisticated techniques to investigate these questions, and the most indispensable is the DNA microarray assay. This apparatus is even more useful than it is colorful, so let's see how this works now.
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Пікірлер: 157

@jamilakalilj 3 жыл бұрын

Currently, using this to supplement my Master's program. The visuals helped to solidify my readings. Thanks, Dave!

@mariemasarikova8575 2 жыл бұрын

This is a quite old comment I know, but wanted to let you know that it's comforting to see that even a master's student needs clarification on this. I'm doing my bachelor's and currently battling imposter syndrome, so thank you.

@m.b.k642 2 жыл бұрын

@@mariemasarikova8575 I’m doing this as a 16 year old , SED

@aahana8816 2 жыл бұрын

@@mariemasarikova8575 same, seeing so many comments from masters students is making me feel better about not being able to grasp the concepts of such techniques easily

@lashazhvania6813 2 жыл бұрын

@@mariemasarikova8575 Haha, thanks for clarifing my syndrome= imposter syndrome-" It disproportionately affects high-achieving people, who find it difficult to accept their accomplishments" I thought i was able to find cure for HIV :D

@robinletourneau3567 2 жыл бұрын

I just wanted to give a heartfelt thank you for this video. My first year final tomorrow is an essay about using microarrays to identify heterochromality genes in huskys and this helped me study for it tremendously.

@giov6538 Жыл бұрын

This is great: super simple, super precise, it makes you really understand the basis of microarrays. From here you can go to more complex videos, but you do need a basis like this to start! Congratulations

@riecchi Жыл бұрын

thank you SO much. i was struggling to understand amidst so much stress but this completely cleared up my confusion and i could not be more grateful right now

@narayanappam6482 3 жыл бұрын

Professor your explanation is informative, precise and beautiful thank you

@bracken7794 2 жыл бұрын

Many thanks for this explanation professor Dave!

@user-le6hp6vr7b 3 жыл бұрын

The explanation was excellent !! i used it to explain the DNA chips for my students

@victoriariosvazquez2924 2 жыл бұрын

Watching this to clarify my mind on my thesis proposal tomorrow, you're a life saver! THX

@asmashakir7859 4 жыл бұрын

Thanku very much professor your video really helped me in understanding microarray the way u explains is best

@camilaglee23 4 жыл бұрын

Thank you very much, you helped me understand much better. Greetings from Argentina!

@thestudymuse_myrsini 3 жыл бұрын

Thank you so much for the effort you put into this, its was so easy to understand

@sainidhi5617 3 жыл бұрын

Amazing video👌👌👌. It was really helpful. Now the concept is clear for me. Thank you soo much for this video 😃.

@semb12 2 жыл бұрын

I'm doing a biochemistry masters and this is helping me - thanks.

@ratethingies6139 3 жыл бұрын

Huge thanks jumping from Saudi to you and the channel!

@krishnaprasad5508 2 жыл бұрын

thank you for your content. you explained it beautifully

@KnighteMinistriez 4 жыл бұрын

I really like how you explain the science. I like these video.

@PoojaSingh-kk6qp 3 жыл бұрын

Thank you sir very beautifully you explain the procedure

@swarnavadutta4648 3 жыл бұрын

you just nailed it🔥 love from India💝

@mongotrip9999 4 жыл бұрын

good work! thanks from austria!

@i.am_pluto4012 3 жыл бұрын

Thank you prof Dave... Much love!!!

@hfof Ай бұрын

omg. You are so talented man, the more I grow up, the more I realize how genius and knowledgeable you are. Cheers to you from Saudi Arabia.

@mustajabalam3794 4 жыл бұрын

Plz make video on validation and QC of flowcytometry and molecular techniques like RFLP ,SSP and SSOP

@roohisakina5742 3 жыл бұрын

Very impressive explanation.

@songthanh896 3 жыл бұрын

Really helpful, Professor!

@atxbee 4 жыл бұрын

thank you for everything you do!! :)

@annielyncy281 3 жыл бұрын

Wonderful presentation and thankyou 👌

@mohamedabdelhamid5771 4 жыл бұрын

Thank you very much , Professor ❤❤❤❤❤❤🙏🙏🙏

@natasharasebotsa7300 Жыл бұрын

i struggled with this for a long time. Thanks a lot

@ThaoLe-kw2ij 2 жыл бұрын

Super helpful! Thank you very much!

@emannadeem3168 2 жыл бұрын

Thank you Professor Dave 😃

@jorgrudiger2894 Жыл бұрын

Thanks. I watched it just for general interest in this subject.

@lilsnaps6881 3 жыл бұрын

you sir have just saved my grade, i salute you o7

@marhabourinboyeva9025 2 жыл бұрын

Thank you so much. This video is really helpful 😃👍

@hamidkiangaikani 4 жыл бұрын

Awesome! Thank you.

@carlosmonf 3 жыл бұрын

Thanks for the info you made it see easy

@ummehabiba7612 Жыл бұрын

great work! so much helpful in my Master's

@Omxralb 2 жыл бұрын

I FREAKING LOVE YOU PROFESSOR DAVE EXPLAINS!!!!!

@Tigerbear1068 10 ай бұрын

Thank you ! This helped me so much !!

@arezoojokar-pn5si 10 ай бұрын

Amazing video. Thank you ❤❤

@lewietlewiet2632 3 жыл бұрын

Why getting a 90€ Biochemistry-Textbook when KZfaq can help you to understand the topic in about 8min? :) Thank you for your content! It helped me a lot! :)

@oldmedstudent1750 2 жыл бұрын

Dude seriously. I'm learning Biochemistry and required to use the Lehninger book that everyone says is so good but I honestly am learning so much more from KZfaq and only refer to the book when I have to. I'm getting along just fine without it.

@user-fn2xc7dz1i 9 ай бұрын

Thanks Professor Dave

@alicelidman6327 3 жыл бұрын

Thank you so much for the video! I have a question. Does the cDNA have to be denatured, like with NaOH or something, so that it can bind to the single-stranded probes in the array?

@Echodonut Жыл бұрын

yes.

@JasonCarlson 4 жыл бұрын

Totally dig your style and material! Go Science and kick some assays..

@amirsh9710 Жыл бұрын

that was so helpful thanks prof😉

@wassmmd8521 3 жыл бұрын

Good work

@alizaalam7624 3 жыл бұрын

thankyouu😌 I never understood microarrays in my class

@Ravezmalazada 3 ай бұрын

Thanks, I hope you make video about RNA microarrays, it will be so completable !

@adamastiti8606 2 жыл бұрын

Thank you prof

@ismaelnaruto30 2 жыл бұрын

Wow, nice video man

@user-rk2fm3bn3u 7 ай бұрын

Thank you professor ❤

@nolifeols 3 жыл бұрын

Could you please make a video explaining RNA sequencing. Thanks.

@sushmanepal1240 2 жыл бұрын

Thank you so much !

@drshreyaagrawal7285 2 жыл бұрын

Amazing !

@DivineBre 2 жыл бұрын

Extremely helpful

@mass_oak 3 жыл бұрын

great content

@Simi.g Жыл бұрын

thank you

@sujis142 2 жыл бұрын

Thankyou sir ✨

@ma.8805 2 жыл бұрын

YOU ARE THE BEST

@rzellereyes1932 3 жыл бұрын

So for microarray, we are only using one cDNA strand which was made from the template, the mRNA. However, there is one thing that isn't clear to me and that is about the RT-PCR. RT-PCR generates double-stranded cDNA right? How come in microarray we are only using the single strand and not double? Is there any intermediate enzyme that was involved?

@nnankegracearikpo2847 Жыл бұрын

Very helpful ❤

@smitshah8074 3 жыл бұрын

thank you sir

@larysavynohradova8845 3 жыл бұрын

отличное видео, наконец все стало понятно))

@kajalgaur3362 3 жыл бұрын

One of best😍

@Dinamite4 2 жыл бұрын

What kind of sequencing can you perform that Dave explains at 7:15? can you do Next gen sequencing?

@huihuihuihuihuihui1 4 жыл бұрын

Nice!

@aimenmustafa62 2 жыл бұрын

Every time Authentic content ...

@ssvr 3 жыл бұрын

Is this this DNA microarray process described in the video also known as "bulk" RNA sequencing?

@moonyqasem6186 Жыл бұрын

Thank you so much ❤ your my hero 🦸‍♂️

@tolerafufa5683 Жыл бұрын

VERY NICE

@detectedoutside8155 4 жыл бұрын

How have you not already made a connect four?

@shahryarkhorasani137 Жыл бұрын

Thank you! One Question: How the RNA splicing during post-transcriptional modification does not cause any problems for RT-PCR?

@IntellicastFacts Жыл бұрын

In a nutshell, RNA splicing involves the removal of introns (non-coding parts of the mRNA) and joins the coding sequences (exons) together in order to enable translation. Since the parts removed are non-coding, it does not affect RT-PCR. RT-PCR, standing for Reverse Transcription Polymerase Chain Reaction, involves reverse transcriptase - converting RNA into cDNA - and (often) Taq DNA polymerase - completing the DNA after denaturation. To sum up, since the RT-PCR process only duplicates CODING DNA, DNA splicing does not affect the process AT ALL, since it removes NON-CODING parts of the original DNA.

@jananishambhasivam7952 4 жыл бұрын

Tku sir I wish I could have been your student.

@sofianbio4997 2 жыл бұрын

Thank you please more things explain about genotype

@magosInformaticus 4 жыл бұрын

I'm a little confused that the approach is to make double-stranded DNA with DNA polymerase and then try and get it to apparently separate and bind to single-stranded probes. Do the probes test for both the sense and antisense strands on each spot?

@samoleulmi990 3 жыл бұрын

Same issue here , couldn't manged to understand that point.

@Behillod 2 жыл бұрын

Hey, you might already have found the answer to your question but in order to make cDNA we use a reverse transcriptase. This is a protein that basically turns your RNA back into DNA. Usually the reverse transcriptase leaves you with a single stranded DNA strand and a round of normal PCR (polymerase chain reaction) is required to turn it into double stranded DNA. Hybridization can then be performed by separating the strands again (either heating or chemical, where heating would be preferred since it is less harmful for the DNA). The stand that is complementary to the probe that was designed will then bind to either one of the strands and start the microarray assay. Which strand binds should not matter too much since the probe should be specific for a certain gene. Hope this helps.

@maylia3132 2 жыл бұрын

this man is a legend i have an assignment due tmrow that if i fail im gonna have a 70 and this man just saved me

@sciencenerd7639 2 жыл бұрын

amazing

@elfa4231 Жыл бұрын

Does this mean that we need to have a full genome sequenced of that specific organism in order to construct the microarray plate?

@thadbonzon 2 ай бұрын

Could someone explain, how do we know where the complementary strands in the microarray came from (as in, from cell type 1 or cell type 2)?

@jonathankimkful 2 жыл бұрын

med student here. wish our professors can teach like this...

@leonr.8084 3 жыл бұрын

Thanks so much, helped a lot for my coming exame!! Greetings from Germany

@meryemlahbara2834 10 ай бұрын

❤❤ thank you

@rowanessam9154 3 жыл бұрын

you are a legend

@azifahahmed4462 3 жыл бұрын

I have a question How could you use Gene Expression Arrays to identify alternative splicing

@calebm9000 3 жыл бұрын

Some researchers have used algorithms to study expression differences between samples. pubmed.ncbi.nlm.nih.gov/11435406/

@mustajabalam3794 4 жыл бұрын

Great

@ananyayadav9686 3 жыл бұрын

This was so amazing...but not gonna lie , the lizard freaked me out a little bit.

@CryptoBountyHunt 2 жыл бұрын

My question is mRNA already bound and complementary cDNA is formed then how they bind with dna probes present in wells

@saroshvachha1588 3 жыл бұрын

100000 times better than my textbook

@berndelignie8002 2 жыл бұрын

Wont you use q pcr for expression?

@julianasuarez7974 3 жыл бұрын

How is the double stranded cDNA "split" into a single strand to hybridize with the probes in the microarray?

@nurajafar8366 2 жыл бұрын

An RNAse endonuclease would have to disassemble the RNA molecule from the cDNA, to allow for the hybridisation

@cdorman11 2 жыл бұрын

Heating, same as for PCR with DNA polymerase

@fidamuhammadkum 4 жыл бұрын

please sir discribed the discovery of drug through micoarray

@tolerafufa5683 Жыл бұрын

YOUR LESSON IS ATTRACTIVE AND GUIDELINE FOR US. BUT I HAVE A COMMENT. HOW CAN I CONTACT YOU PERSONALLY?

@jaykay8098 2 жыл бұрын

word of caution, rna pol doesnt need a primer right, so reverse transcriptase having a primer feels fishy but its not because although widely called reverse trasncriptase the enzyme itself is synthesizing dna and so in essence its a dna pol and dna pols need the TTT primers.

@polymath5093 2 жыл бұрын

I have a question here.. Do we amplify cDNA before introducing it to the Array? If yes, we already used a primer to amplify our gene of interest.. In this case, how different genes in the array well would produce color except the one we amplified with our primer? If we don't amplify, how the trace amount of different cDNAs will produce color in array?

@prabhar4254 10 ай бұрын

Actually..i had the same doubt b4...but now i got to know that if we are adding gene specific primer means, no need to go with microarray since it is a highly powerful tool for differentially expressed genes not for gene expression, i think...im not sure about it.....if you know something means, please leme know bcx iam having a seminar on bee nutrugenin ICS..

@caddyshackwizard9367 4 жыл бұрын

Is it me or does it seem like Dave's videos are being suppressed the amount of views to subscribers just does not add up

@ProfessorDaveExplains 4 жыл бұрын

Nah it's just that people subscribe for so many different subjects, any given video will only appeal to a small percentage of them. But spread the word!

@adnanqamar7815 2 жыл бұрын

Genius

@vasundhararaina6849 3 жыл бұрын

so you mean a common pool with all the different samples is allowed to hybridise on the array

@clashwithsparkcrazygamers Жыл бұрын

During the process of RT-PCR we get a complimentary ssDNA for which we perform PCR to get a dsDNA, we add this to each of the wells of the array plate and the complimentary DNA probe hybridizes and form a link. My doubt is if the DNA is already ds post RT-PCR how will it bind to another DNA probe. Or are we considering the DNA to be ss post RT- PCR. (the animation does not support the fact that the DNA is ss post RT-PCR at 5:11 ) Anybody?

@swiya8577 10 күн бұрын

same question

@chocolatecrud 3 жыл бұрын

yup

@sriram-ug2ly 2 жыл бұрын

Sir but how can this be used for quantitative assay of a particular gene...i mean i only understand that this will be only useful for qualitative analysis

@orange123yoshi 2 жыл бұрын

the general idea: with each pcr cycle, the amount of a certain DNA sequence is double. Using some kind of labeling, e.g. fluorescent dyes, we can detect the presence of the mentioned DNA sequcense. The fluorescent dye has a detection limit (minimal concentration of the substance which can be detected), so, if we know that we detected the DNA after, let's say, 9 PCR cycles, and the detection limit for the dye is 100 nmol/ml, that means that the amount of DNA in the initial sample was 2^9 times lower, thus, quantitative analysis.