Primer designing for real time PCR using NCBI Primer Blast 

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.

Thank you so much! Once I returned home from the lecture, I realized that I do not remember shit.

Thank you very much. It was beautifully explained.

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.

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 :)

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?

very good video for people!

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

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

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

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

Good information

How can I record laptop monitor that I design primer ?

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.

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

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

thanks really helpful

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

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?

nice explanation...

How to do primer sequencing in plant gene

Lovely, thanks

Very nice explanation. Thank you :)

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

Sir some problem please help me RT PCR primer design

sir, i want to know about entrez query

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

Are you malayali

HELLO SIR ITS TAKING TOO MUCH TIME TO GET THE RESULT CAN YOU PLEASE HELP TO SOLVE THIS ISSUE

Dankeschön

Hello sir.. I am trying to make primer for plant sample. Itried to do as shown in the video but it is showing error like. The primer specificity can not be determined as sequences for the selected organism are not present in selected database: refseq_mrna please help me with this.

Thanks!(:

Great

Lee Frank Thomas Patricia Perez Dorothy

Hello , thanks for your explanations . I wondered if i can ask some questions . Iam a total newbie in PCR . I used Perlprimer app for making primer for CFTR gene but the result in NCBI site for my primer was different with the app eventhough i considered the same situation for both can u please help ? 🩵