Primer Design for PCR

  Рет қаралды 660,343

Herbert Sauro

Herbert Sauro

Күн бұрын

Пікірлер: 139
@alisonlee1891
@alisonlee1891 9 жыл бұрын
Thank you so much for this!! I looked everywhere on the internet and for some reason, the way you worded it specifically and drew everything out just made so much sense to me!! Thank you!!
@إبراهيماشرف-ل3و
@إبراهيماشرف-ل3و 3 жыл бұрын
Could you believe it, me too!
@FeraleHubbard
@FeraleHubbard 4 жыл бұрын
You. Are. Awesome. You just explained something that two profs and 2 TA's failed to be able to explain to me in such an understandable way. Thank you!!
@KrishnaBhat1988
@KrishnaBhat1988 10 жыл бұрын
Nice explanation, but one suggestion when you compare sequences, try to put the video frame in such a way that both the sequences are seen at a time.
@MaideCanyakar
@MaideCanyakar 5 жыл бұрын
I agree
@qunyangjiang5360
@qunyangjiang5360 Ай бұрын
Very nice presentation to tell me the determination of F and R primers!!!! Thank you very much!
@cayla489
@cayla489 10 жыл бұрын
Great video! My only comment would be to be a bit more aware of what can be seen on screen at a given time, as sometimes things were a bit cut off at the edges.
@daniellebelize1862
@daniellebelize1862 9 жыл бұрын
Thanks, it's been a while since I had anything to do with primer design. It was fun to relive it.
@andreaspurnomo1688
@andreaspurnomo1688 3 жыл бұрын
I'm digging for gold but found diamond instead, 8 years late, thanks for the videos!
@wendahu5943
@wendahu5943 4 жыл бұрын
It's nice explanation. FYI: I understand it's for better explanation, but the template you choose is quite short isn't it, so that primers actually overlap. I doubt these primers will work because they form dimers too easily.
@goldmanuniversities
@goldmanuniversities 5 жыл бұрын
great work done sir but i recommend if you can edit to make both strands visible at 9:40
@alexcracker
@alexcracker 5 жыл бұрын
At 11:50 you say :"This low temperature annealing G and C means that when the temperature is increased in PCR this primer physically can not bind...." It is not correct. The primer with higher GC-content required higher melting temperature.
@alextzarax3865
@alextzarax3865 5 жыл бұрын
True
@joannathomas5380
@joannathomas5380 10 жыл бұрын
Shame that the bottom of the screen is cut off
@yimuyang4101
@yimuyang4101 9 жыл бұрын
I m more addicted to the gentle voice than the helpful video lol thanks!
@ailshajoya5760
@ailshajoya5760 5 жыл бұрын
i still confused about the primer forward and the primer reverse,anybody can explain to me in simple way? thankyou 😔
@juggernaut407
@juggernaut407 2 жыл бұрын
I'm abit confused. How many nucleotides would be in his forward primer?
@muhammadtufail3323
@muhammadtufail3323 4 жыл бұрын
excuse me will u plz tell me that if i have a DNA sequence of 3819 bp and i want to make primers from this sequence then can i start from anywhere in the sequence or just for fwd primer i would have to start from begining and for reverse i would start from the end of the sequence??
@TonyTigerTonyTiger
@TonyTigerTonyTiger 3 жыл бұрын
1:00 "What I've drawn down here" Where? Can't see it.
@Athams
@Athams 10 жыл бұрын
I want to ask, why is 3 prime and 5 prime standard convention? Is there a video where I can check this out?
@jskratnyarlathotep8411
@jskratnyarlathotep8411 4 жыл бұрын
Where could I find explanation on what happens when primer binds to a different region of the plasmid with exact nucleotide sequence? How then that regions not get amplified to the comparable amount as the region of interest?
@stephenrose1902
@stephenrose1902 6 жыл бұрын
I'm confused, if the DNA polymerase binds to the 3' end and goes in the three prime direction, there's no where to go, its already at the end?
@tatietatty
@tatietatty 6 жыл бұрын
Hi, it is about SSR marker. how may I know if the ssr has most fixation and similar number of genetic fingerprint?
@shailenderrajput3500
@shailenderrajput3500 4 жыл бұрын
How this RVs can attach ..since it is changed ????
@jonathanaltamirano6915
@jonathanaltamirano6915 5 жыл бұрын
Are you saying that you have to reverse compliment the reverse primer or that you just have to reverse the sequence of the reverse primer? I'm asking because when I put in the original reverse primer that's in the 3'-5' orientation into a reverse compliment calculator I get the complimented sequence of your reverse primer and not the sequence you say is the reverse compliment.
@joedart1465
@joedart1465 5 жыл бұрын
If you turned the molecule upside down so that the top strand was the bottom strand, then you could read that strand left to right correctly. The problem is the way we lay the molecule on the page. The bases are complementary but because the molecule is laid out like that we have to read the base sequence backwards to make it forward. notice that the forward primer can be read forward simply because we are reading it left to right.
@missjade2940
@missjade2940 Жыл бұрын
Hello Herbert, First off, thanks for doing this tutorial, great help to me. I wanted to ask, why not just make it 18 base pairs because there's already a G assigned on the 18th position. Is there a reason for not doing the suggested?
@iya3952
@iya3952 4 жыл бұрын
Can anyone tell me how they actually synthesize the primers??
@Theprofessionalsurgeon
@Theprofessionalsurgeon 3 жыл бұрын
dm me
@iya3952
@iya3952 3 жыл бұрын
@@Theprofessionalsurgeon i can’t dm through here
@harshaakariyawasam6322
@harshaakariyawasam6322 3 жыл бұрын
Thanks a bunch for this simple and brief explanation. It was really interesting! Keep it up!
@naderanoori373
@naderanoori373 7 жыл бұрын
thank you so much, actually you have save my many hours of studying, to get to learn how to design a primer.. keep it up
@umarecol81
@umarecol81 8 жыл бұрын
i stil don't understand the fact that we have to reverse the primer, I mean it would still come out the same?
@Dreamslayer0987
@Dreamslayer0987 8 жыл бұрын
It's simply when you are buying and ordering your primers from somewhere. Because the structure will be different if you go from 3' to 5' and 5' to 3' and if you ordered it as 3' to 5', you'll be getting a 5' to 3' primer from the vendor and it won't work as you imagined it to do.
@MrRyanWonderlin
@MrRyanWonderlin 7 жыл бұрын
Do you need to include your start codon in your forward primer?
@MrRyanWonderlin
@MrRyanWonderlin 7 жыл бұрын
What about the stop codon in the reverse?
@doraliciacasaresdelarosa273
@doraliciacasaresdelarosa273 7 жыл бұрын
What do you need a start or a stop codon for in a PCR?
@joedart1465
@joedart1465 5 жыл бұрын
the forward primer is what initiates the polymerase.
@sophie4153
@sophie4153 7 жыл бұрын
stupid question: when does the DNA pol. stop copying the strand? How many bp from the primer?
@sophie4153
@sophie4153 7 жыл бұрын
Wait wait wait... just where the two primers end, so you get dsDNA between the primers?
@abelbabel8484
@abelbabel8484 6 жыл бұрын
Polymerase stops wherever the template ends
@vivianegirardin5667
@vivianegirardin5667 9 жыл бұрын
Very easy to follow and well explained, thank you!
@debbiejoyable
@debbiejoyable 4 жыл бұрын
What software is being used for the illustration?
@kaleemullahmarwat1207
@kaleemullahmarwat1207 3 жыл бұрын
Why we need partial sequences while designing primer
@Itsmesvlogerr
@Itsmesvlogerr 6 жыл бұрын
sir, both primer are complementary to each other like dsDNA?
@joedart1465
@joedart1465 5 жыл бұрын
No. The primers go to the 5 prime ends of the two strands which are not complementary. (they could be but that would be coincidence.
@cloudy7817
@cloudy7817 14 күн бұрын
thank youuuuuu🙏 😭-a struggling brandeis student
@shameeramahawattage7150
@shameeramahawattage7150 4 жыл бұрын
Great Video. Short and really understanding
@GrammeStudio
@GrammeStudio 7 жыл бұрын
what are sticky ends for?
@joedart1465
@joedart1465 5 жыл бұрын
If you want to splice two pieces of DNA to each other you can do it by stripping away a few bases from the 5' ends of both (assuming they are complementary ) and they will tend to ligate with a little help.
@aregawiataklti2177
@aregawiataklti2177 8 жыл бұрын
please could you help me on primer design
@ahmedalshal97
@ahmedalshal97 Жыл бұрын
Thank you for the detailed explanation
@sharmilasrinivasan9417
@sharmilasrinivasan9417 3 жыл бұрын
Just what I was looking for!
@afnanzuhdy4470
@afnanzuhdy4470 3 жыл бұрын
Great video, simple explanation, thank you for this man!
@sudhirsawarkar4575
@sudhirsawarkar4575 8 жыл бұрын
Simple and clear explanation ... thank you
@claire2021
@claire2021 7 жыл бұрын
Thank you!!! I understood it perfectly. And guys, If you have SOME idea of basic genetics you wont need the view of the whole screen... Greetings from argentina!
@dr.neetisharma6546
@dr.neetisharma6546 4 жыл бұрын
Thank you so much for this video. Nice Explanation. So much helpful.
@3923-x1e
@3923-x1e 8 жыл бұрын
Why do you have to have 20/21 bases for the forward primer, must you? Or is it just desirable?
@icelysis3974
@icelysis3974 8 жыл бұрын
Found this on the internet: Primer Length: It is generally accepted that the optimal length of PCR primers is 18-22 bp. This length is long enough for adequate specificity and short enough for primers to bind easily to the template at the annealing temperature. Hope that helps!
@waterloochavez3531
@waterloochavez3531 7 жыл бұрын
also it would be less likely for there an equal sequence if the length is greater than 17-18 bp
@dr.domingouez3856
@dr.domingouez3856 Жыл бұрын
يعطيك العافية
@terrylaitw840
@terrylaitw840 8 жыл бұрын
very useful and informative. Great job there
@queseraseraozi
@queseraseraozi 10 жыл бұрын
Wonderful work my friend , please keep doing this ! Regards
@aaronledray4505
@aaronledray4505 9 жыл бұрын
Thanks! Great refresher.
@sumana301
@sumana301 10 жыл бұрын
thank you!! I was so confused until now. THANK YOU
@ABDULKHALIQ-uw8bn
@ABDULKHALIQ-uw8bn 5 жыл бұрын
NOWadays i am sure you have confusion, but i have a lot of confusion PLEASE guide me
@natalygg7332
@natalygg7332 8 жыл бұрын
Thank you for the precise explanation! You really helped me
@haifamansour24
@haifamansour24 9 жыл бұрын
very simple and clear explanation.. Thanks
@JyotiMishra-pn2eq
@JyotiMishra-pn2eq 9 жыл бұрын
bt hw can i download it
@mohab.m.metwally
@mohab.m.metwally 9 жыл бұрын
+Jyoti Mishra www.google.com.eg/webhp?sourceid=chrome-instant&ion=1&espv=2&ie=UTF-8#q=youtue+downloader+online
@haifamansour24
@haifamansour24 8 жыл бұрын
write ''ss'' before ''youtube'' in the link.
@mohab.m.metwally
@mohab.m.metwally 8 жыл бұрын
Haifa Mansour that is a good website
@MrMetalHead1100
@MrMetalHead1100 7 жыл бұрын
Thanks bro. That was everything I needed!
@minotigupta6714
@minotigupta6714 3 жыл бұрын
Thank you sir ☺️this video was very helpful 😊
@sallycha7952
@sallycha7952 6 жыл бұрын
Best explanation! Thank you so much!
@jamshedfbc
@jamshedfbc 5 жыл бұрын
Excellent video with poor camera frames. Overall, good effort. Liked already.
@ThunderChunky101
@ThunderChunky101 4 жыл бұрын
Would it be weird if you found an identical 18 nucleotide long primer in the DNA of a virus and a human? Anyone have a clue if this is common?
@muhammadtufail3323
@muhammadtufail3323 4 жыл бұрын
kindly reply me asap becoz it is very important for me to know that...
@kimp.dr.n2652
@kimp.dr.n2652 2 жыл бұрын
Can't see the whole video.
@Guzinator555
@Guzinator555 6 жыл бұрын
play it on 1.5 speed, better to follow
@mariyaseb
@mariyaseb 11 жыл бұрын
Very good video...
@neelparmar6690
@neelparmar6690 7 жыл бұрын
Fantastic vid, thanks so much
@心木
@心木 7 ай бұрын
Very nice
@9gatosem2010
@9gatosem2010 11 жыл бұрын
excelente
@linusbao8650
@linusbao8650 6 жыл бұрын
thank you so much, very helpful
@为欢
@为欢 4 жыл бұрын
very good video
@vishwambarnavale5740
@vishwambarnavale5740 7 жыл бұрын
It's really helpful
@desysamsung8729
@desysamsung8729 6 жыл бұрын
Great job!
@CallingtoAllahSWT
@CallingtoAllahSWT 8 жыл бұрын
Very helpful. Thank You
@TheZumana
@TheZumana 11 жыл бұрын
very well explained
@panijani
@panijani 6 жыл бұрын
Thank you for the video!! I learned something ;)
@julles81981
@julles81981 11 жыл бұрын
This is a great video thank you so much
@lemotheemonemo
@lemotheemonemo 9 жыл бұрын
Thank you for this. It was very helpful! ^_^
@marlamiranda5900
@marlamiranda5900 7 жыл бұрын
Very helpfulll, thank you so much !!!
@AyazSamo
@AyazSamo 5 жыл бұрын
Marla Miranda when’re are you? Marlo
@debabratadutta6
@debabratadutta6 4 жыл бұрын
Thanks
@bh5169
@bh5169 2 жыл бұрын
thank you!
@zenamahrab2873
@zenamahrab2873 5 жыл бұрын
هل من مترجم؟؟
@thortagro
@thortagro 5 жыл бұрын
So much helpful. Thank you very much man:-)
@mirellanavarreteflores1134
@mirellanavarreteflores1134 7 жыл бұрын
Wow, thanks amazing video
@nt593
@nt593 7 жыл бұрын
Great , thank you very much
@TheScienceSpot724
@TheScienceSpot724 4 жыл бұрын
thanks a lot
@havenkerley176
@havenkerley176 6 жыл бұрын
thank you
@beautyp.9069
@beautyp.9069 6 жыл бұрын
Very helpful!
@km2052
@km2052 5 жыл бұрын
thx
@nourghaddar4298
@nourghaddar4298 8 жыл бұрын
Thank you amazing video !
@melon8419
@melon8419 6 жыл бұрын
very helpful! thx very much
@km2052
@km2052 5 жыл бұрын
SYNTHETIC biology , thx
@phagyauto160
@phagyauto160 6 жыл бұрын
太有用了
@nazimmedzhidov9257
@nazimmedzhidov9257 11 жыл бұрын
Thank you for this video!!!
@mjresina
@mjresina 6 жыл бұрын
So useful
@yaraedor2881
@yaraedor2881 6 жыл бұрын
THANK YOU SO MUCH
@frankaddeo6132
@frankaddeo6132 7 жыл бұрын
thanks a lot!
@Running99fly
@Running99fly 9 жыл бұрын
thanks.
@green111111111
@green111111111 6 жыл бұрын
thank you soo much!
@AmruMagdy
@AmruMagdy Жыл бұрын
نريد الجلوس بجوار النبي صلى الله عليه وسلم في الجنة ....
@alukooluseun8661
@alukooluseun8661 4 жыл бұрын
NOW AM INTERESTED IN THAT VIDEO BUT COULD NOT
@marygracesedanza6085
@marygracesedanza6085 10 жыл бұрын
THANKS i learned many things! can someone help me go about designing a degenerate primer for aquatic species?i would greatly appreciate it
@froilananthony7391
@froilananthony7391 4 жыл бұрын
sars cov2 primer
@nicolecrosby7530
@nicolecrosby7530 11 жыл бұрын
good one!thank you)
@VoxMortui
@VoxMortui 5 жыл бұрын
This helped so much, thank you!!
@ghostmedic86
@ghostmedic86 5 жыл бұрын
This video would have been significantly more helpful if we could have seen everything on the screen you were pointing to. On our end, your pointing and talking about something that is a couple inches from or visible screen. Thanks anyways though.
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