Biotechniques | Principles of Primer Design for Full Gene Amplification

Well understood. Thank-you

This is exactly what i needed, thank you so much

Thanks 😊

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?

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

Where are the links ????

This is very helpful! Thank youu!

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

Why Forward primer sequence remains same as complementary strand ?

Your vids are concise and very simple to understand

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

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

very helpful, thank you !

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

Thanks a lot❤

Thank you!

Why we need partial sequences whlie designing of primer?

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

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%)

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

I am so happy I found you!!

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

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?

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

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.

GOOD LECTURE 👍GOD BLESS YOU TEACHER 😊

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

Can we make the primer for whole gene for qPCR

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

Really good explanation!!
Helped a lot.

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

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!

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?

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

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

Hey can I send you a message ?

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.

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

Where you brought these genes first?

what

Why cant we get people like this in universitys?

Thank you very much!

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

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

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?

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

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

It is very useful for me , thank you so much