Primer designing for real time PCR using NCBI Primer Blast

This video explains how to design primers for real time pcr using primer blast.
NCBI's primer blast tool helps in designing primers and also allows to check the specificity of the designed primers.
Various options are available to improve the primer stringency.

[annavl]

Hi, thanks for the video! Another way I know to avoid potenitial genomic DNA amplification is by selecting "Primer must spand the exon-exon junction", so no intron-including DNA would be amplified.

[krishnaprasad]

Thank you very much. It was beautifully explained.

[Scienceiirwn]

Amazing Video!!! Is there anyway you can do a video on designing a primer to target different variants ? for example i noticed in your example there are many GAPDH variants (V1 -V4), what if you want to target the different variants to see the expression of each one?
I would really appreciate if you can give an example using CD44 which has 10 variants.

[anuranjanapv]

Hello sir, thank you for the video!
How to select the variant of our gene of interest? On what basis we have to choose?!

[ayeshamalik]

Hi, thanks for this wonderful Video. I learnt a lot from your videos. Can you please make a video on how to analyze sanger sequencing through Geneious Software and MEGA? Will be much Appreciated. Thanks

[somayehmohammadi]

Hello .How to be extract PSSM profiles and to be generates matrix? I have consider linear substitution probability matrix.

[hbBilzZ]

Thanks for the video, it was very helpful. Is the process the same for qPCR primers?

[sanjuktaghosh]

Hi, a very useful video. Could you link the video in which you have discussed how to increase stringency so that the primers become more specific. Kindly reply :)

[anishamn]

How you assign a particular value to Tm. What is the criteria for that value?

[papajiang]

I am wondering how does this tool guarantee the primers are specific to human? Can I say once you input the organism option as human, then the tool will help you to design specific primers to human? And if the primers only have one SNP to a different species, does it still call specificity just due to the one SNP?

[mohammadrezasamiee]

Hi thanks for the video, but I didn't see probe for real time pcr

[murchanabarua]

I have a question? Using this website how would I make a forward and reverse primer that is 18-35 nucleitodes long? because whenever I put that into "PCR product size" from min 18 to max 35 it doesn't work.

[AZMERANDA]

Thank you for the video, my question is, what I should do if the primers were not found? Is there another way to design primers?

[bijayininayak]

How to do primer sequencing in plant gene

[beinghuman]

sir i have a confusion ?
when we have to select nucleotide and when we have to select gene in the database ?

[egamberdimaxmudov]

How can I record laptop monitor that I design primer ?

[chinonyeonyeulo]

I want to run a primer blast for a microorganisms, how do I run the DNA sequence?

[yunasali]

sir, i want to know about entrez query

[prabhubiograd]

The video must have included probe design too in case of real-time PCR

[biologywithboss]

Sir some problem please help me RT PCR primer design