THANK YOU! I have a project where i need to design a primer and your video genuinely saved me and explained it suuuper well too!

FINALLY!. Exactly what I was looking for! Thank you!

This is the best explanation I have come across!! Thanks

Very informative. I am a beginner and this is what I was looking for. Thanks.

Wow! this is just the answer to the questions I have been trying to ask. THank YOu

The video is easy to comprehend and implement in primer design. Thank you for a job well done

Thank you! Great explaination! I have to do this for my homework and this is the first video that really helps! Thanks

This is exactly what i needed, thank you so much

Thank you so much your perfect lesson of primer design!!

I am so happy I found you!!

Really good explanation!! Helped a lot.

GOOD LECTURE 👍GOD BLESS YOU TEACHER 😊

Thank you so much for your video. I am wondering how we check the accuracy of the design primer. Is there any website like you mentioned in the video? I wanna see if my design has appropriate Tm, and GC content and no secondary branching.

From here you would have to deterimine the Tm for each and hope that they are close to each other for the PCR to work.

This is very helpful! Thank youu!

Your vids are concise and very simple to understand

its very helping to me.. i was looking for target gene forward and reverse primers form.published literature but it was lacking number of nucleotides and qPCR protocol for ea ch target gene.. the published literature only contain gene accession code where the origin gene sequenceing section can be obtained .. bu using available information in literature i could be able to design primer by learning this tutorial.. Thank u for sharing this Knowledge

Well understood. Thank-you

Many thanks for video. 1. Primers you designed, how to get to know the annealing temperature of both primers and GC . 2. What is difference between full length primers and short length primers means why we do. For example one gene express in full length primer in semi-qPCR but in short length designed primer this isn't expressing.

It is very useful for me , thank you so much

very helpful, thank you !

Thanks, that was helpful

Nice explanation, you helped me alot with my homework. Thanks very much!

THANK YOU LIFE SAVER

I could not find the description table I need the websites, please

Thank you very much!

but how would you check for dimer formation ?

Maybe add these links for sites in the description below of the video?

Hi, what app did you use to make the video - with the coloured pens on the right? What tool is this, please?

Thank you!

Thank you for this awesome vidoe. Can you please tell.me how can we check the orientation of DNA sequence. Please help

Hey there! Thanks for the video. What if the forward primer as described in the video contains GC and AT rich regions which are not desired? Or if they have suboptimal melting temperatures? Would one then have to search for other 20mers upstream and downstream of the gene to use instead of the start and stop regions as starting points for the 20mers? And how would we do that? Is there a tool for that also? Kind regards

Thank you so much, perfectly to the point. would you also please make a video on how to design a primer for SNP genotyping? or is it the same?!

Can you put out some practice questions for us? Thank you

Thanks a lot❤

The link isnt working for me? Has anyone else had success and can post the link?

Thank you great video. But does your fp need reverse complement as well? Thxxx

we need to add the restriction site before the forward primer and reverse primer and we also need to add the stuffner nucleotides before the restriction site but do you have nay idea how many base pair we can add before the restriction site?

What about the quality of the primers are they any good in terms of the Tm and self complementary?

Sir I like your video I want to confirm that reverse primer is 3 to 5 direction or 5 to 3 direction please tell me?

Can we make the primer for whole gene for qPCR

Your reverse primer doesn't end with a C or a G. Would it work better if it was one base longer to get a better "3' clamp"?

Sir reverse primer is 3 to 5 direction or 5 to 3 direction please tell me the answer?

Hi, in the second method the forward primer is same as template sequence, it should be complementary if it has to go and bind to template strand, how it's gonna work?

Why we need partial sequences whlie designing of primer?

Why use complementary sequence for RP but not for FP..??

Why Forward primer sequence remains same as complementary strand ?

Where are the links ????

But how will you check if there are off-target amplification?

perfect

Thanks 😊

I don't understand what the "optimized" primers are. What are they optimized FOR?

I just did my question paper with your explanation if I don't get marks I m going to hunt you down 😂😛

Does this apply for RT-PCR as well? Template is an RNA virus.

Where you brought these genes first?

In the video why forward and reverse primer has same 5 prime to 3 prime direction

Great, but did not explain why the Primer -Blast did not work and you had to use another tool? also why did you not specifiy amplification ranges in the primer blast?

how to design probe

How about a link to those websites? Wouldn't that be useful...

for the reverse primer, why did you not change the 5 prime to 3 prime and 3 prime to 5 prime.i actually thought you forgot to do that.

But amplification of large gene with only one primer isn't trustworthy or may have incorrect base sequence

Why cant we get people like this in universitys?

When they say "it lacks the primer for reverse transcriptase" ... uhm, it can make its own!

Hey can I send you a message ?

The start-point and endpoint of a desired DNA-sequence does not provide necessarily a suitable set of primers. In most cases, it does not. (Selfdimers, wrong amount of Cytosines en Guanines. CG-amount should be around 60%)

what

Your explanation for the forward primer makes no sense tbh. How is that primer going to attach to the first 20 base pair sequences of that strand when it's the exact same and isn't a reverse compliment?

Sir I wanna a contact you I need you help ...