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.
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.
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!
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.
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?
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?
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.
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??
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.
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!
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!
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.
I m more addicted to the gentle voice than the helpful video lol thanks!
great work done sir
but i recommend if you can edit to make both strands visible at 9:40
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
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!
Shame that the bottom of the screen is cut off
Just what I was looking for!
Wonderful work my friend , please keep doing this !
Regards
Thank you for the detailed explanation
Great Video. Short and really understanding
Thanks! Great refresher.
Thank you so much for this video. Nice Explanation. So much helpful.
i still confused about the primer forward and the primer reverse,anybody can explain to me in simple way? thankyou 😔
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
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
Great video, simple explanation, thank you for this man!
Simple and clear explanation ... thank you
Thank you for the precise explanation! You really helped me
Best explanation!
Thank you so much!
very useful and informative. Great job there
Thanks bro. That was everything I needed!
Very good video...
Thank you sir ☺️this video was very helpful 😊
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.
Excellent video with poor camera frames. Overall, good effort. Liked already.
Fantastic vid, thanks so much
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?
Very helpful. Thank You
thank you so much, very helpful
Thank you for this. It was very helpful! ^_^
It's really helpful
Great job!
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?
Thank you for the video!! I learned something ;)
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?
I want to ask, why is 3 prime and 5 prime standard convention? Is there a video where I can check this out?
Wow, thanks amazing video
So much helpful. Thank you very much man:-)
Great , thank you very much
I'm abit confused. How many nucleotides would be in his forward primer?
Very helpful!
This is a great video thank you so much
Thank you amazing video !
Hi, it is about SSR marker. how may I know if the ssr has most fixation and similar number of genetic fingerprint?
very helpful! thx very much
very well explained
Very helpfulll, thank you so much !!!
Marla Miranda when’re are you? Marlo
يعطيك العافية
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.
Why we need partial sequences while designing primer
Can anyone tell me how they actually synthesize the primers??
dm me
@@Theprofessionalsurgeon i can’t dm through here
Very nice
Thank you for this video!!!
very good video
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??
1:00 "What I've drawn down here" Where? Can't see it.
How this RVs can attach ..since it is changed ????
This helped so much, thank you!!
So useful
excelente
What software is being used for the illustration?
THANK YOU SO MUCH
thank you soo much!
thank you!
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
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.
good one!thank you)
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.
SYNTHETIC biology , thx
please could you help me on primer design
thanks a lot!
play it on 1.5 speed, better to follow
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.
THANKS SO MUCH!!!!!
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
thanks a lot
thank you
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?
Can't see the whole video.
kindly reply me asap becoz it is very important for me to know that...
太有用了
NOW AM INTERESTED IN THAT VIDEO BUT COULD NOT
Thanks
THANKS i learned many things! can someone help me go about designing a degenerate primer for aquatic species?i would greatly appreciate it
thanks.
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.
thx
هل من مترجم؟؟
نريد الجلوس بجوار النبي صلى الله عليه وسلم في الجنة ....
sars cov2 primer
DP lol