Biotechniques | Principles of Primer Design for Full Gene Amplification

In this video, I will show you how to design primers to amplify the entire gene during a routine PCR.

[CarolinaVarotto]

Thank you so much! I'm involved in a project where they asked me to design primers for the first time and your detailed explanation saved me, thank you again for your work and for sharing your experience with us!

[yipeerika]

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!

[polarisgemini]

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

[joklbeatrice]

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

[MAYOWAANIFATOLASOPE]

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

[jonathandavid]

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

[sunnetinternationalbusines]

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

[alanhassall]

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.

[hiramaryam]

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.

[umairameer]

Why Forward primer sequence remains same as complementary strand ?

[tahirm]

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

[smmbadawy]

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

[funnygov]

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

[alanhassall]

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"?

[sowmyahh]

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?

[KoalaKid]

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

[smartmail]

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

[nandani]

And what will be the amplicon length for the amplified product after PCR?

[janggeumseo]

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

[stephenchapman]

Great video, thank you! Buto use say from a DNA sequence, but the sequence you use as an example is an mRNA sequence