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!!
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!!
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.
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.
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!
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.
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??
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?
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.
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.
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.
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?
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!
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.
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.
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.
I agree
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!!
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!!
Could you believe it, me too!
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.
Thanks, it's been a while since I had anything to do with primer design. It was fun to relive it.
I'm digging for gold but found diamond instead, 8 years late, thanks for the videos!
I m more addicted to the gentle voice than the helpful video lol thanks!
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.
great work done sir
but i recommend if you can edit to make both strands visible at 9:40
i still confused about the primer forward and the primer reverse,anybody can explain to me in simple way? thankyou 😔
Shame that the bottom of the screen is cut off
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!
Thanks a bunch for this simple and brief explanation. It was really interesting! Keep it up!
Very easy to follow and well explained, thank you!
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
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.
True
I'm abit confused. How many nucleotides would be in his forward primer?
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??
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?
Hi, it is about SSR marker. how may I know if the ssr has most fixation and similar number of genetic fingerprint?
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?
I want to ask, why is 3 prime and 5 prime standard convention? Is there a video where I can check this out?
Great Video. Short and really understanding
Thank you so much for this video. Nice Explanation. So much helpful.
Thank you for the detailed explanation
Great video, simple explanation, thank you for this man!
Just what I was looking for!
i stil don't understand the fact that we have to reverse the primer, I mean it would still come out the same?
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.
Thank you for the precise explanation! You really helped me
What software is being used for the illustration?
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.
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.
Simple and clear explanation ... thank you
Wonderful work my friend , please keep doing this !
Regards
thank you!! I was so confused until now. THANK YOU
NOWadays i am sure you have confusion, but i have a lot of confusion PLEASE guide me
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?
1:00 "What I've drawn down here" Where? Can't see it.
Thanks! Great refresher.
very simple and clear explanation.. Thanks
bt hw can i download it
+Jyoti Mishra www.google.com.eg/webhp?sourceid=chrome-instant&ion=1&espv=2&ie=UTF-8#q=youtue+downloader+online
write ''ss'' before ''youtube'' in the link.
Haifa Mansour that is a good website
How this RVs can attach ..since it is changed ????
Best explanation!
Thank you so much!
Thanks bro. That was everything I needed!
very useful and informative. Great job there
stupid question: when does the DNA pol. stop copying the strand? How many bp from the primer?
Wait wait wait... just where the two primers end, so you get dsDNA between the primers?
Polymerase stops wherever the template ends
Can anyone tell me how they actually synthesize the primers??
dm me
@@Theprofessionalsurgeon i can’t dm through here
Why do you have to have 20/21 bases for the forward primer, must you? Or is it just desirable?
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!
also it would be less likely for there an equal sequence if the length is greater than 17-18 bp
Thank you sir ☺️this video was very helpful 😊
sir, both primer are complementary to each other like dsDNA?
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.
Why we need partial sequences while designing primer
Thank you for this. It was very helpful! ^_^
Fantastic vid, thanks so much
thank you so much, very helpful
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?
This is a great video thank you so much
please could you help me on primer design
It's really helpful
Do you need to include your start codon in your forward primer?
What about the stop codon in the reverse?
What do you need a start or a stop codon for in a PCR?
the forward primer is what initiates the polymerase.
Very helpful. Thank You
Thank you for the video!! I learned something ;)
Very nice
يعطيك العافية
So much helpful. Thank you very much man:-)
Very good video...
Excellent video with poor camera frames. Overall, good effort. Liked already.
Great job!
Wow, thanks amazing video
very well explained
Thank you amazing video !
Very helpfulll, thank you so much !!!
Marla Miranda when’re are you? Marlo
thank you!
Great , thank you very much
Very helpful!
very helpful! thx very much
what are sticky ends for?
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.
This helped so much, thank you!!
Thank you for this video!!!
Can't see the whole video.
very good video
thanks a lot
So useful
excelente
play it on 1.5 speed, better to follow
Thanks
THANK YOU SO MUCH
thank you soo much!
kindly reply me asap becoz it is very important for me to know that...
SYNTHETIC biology , thx
thank you
thanks a lot!
good one!thank you)
太有用了
thanks.
thx
THANKS SO MUCH!!!!!
THANKS i learned many things! can someone help me go about designing a degenerate primer for aquatic species?i would greatly appreciate it
نريد الجلوس بجوار النبي صلى الله عليه وسلم في الجنة ....
NOW AM INTERESTED IN THAT VIDEO BUT COULD NOT
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.
هل من مترجم؟؟
sars cov2 primer
wow you're not indian, how can this be
spagetti001 😄