The Features Of A Good qPCR Primer Pair

[@nunitchagucci3047]

This is so wonderful! Save my time of struggling in grad school!
Thank you so much!!! :)

[@jacelinesanches1956]

Can you make video about plasmid construction

[@calebm9000]

Thanks for the video. If using gene-specific primers for quantification/detection, is it as necessary to bind towards the 3' end?

[@vitaliivedernikov4961]

Actually, melting temperature is when 50% of a primer molecules is annealed to the target sequence, not just a half of it sequence!

[@StevenBradburn]

Hi Vitaly. You are completely right. I realised this after I uploaded the video so it is not easy for me to edit the video to correct this.

[@NabiShah]

hi there, could you suggest which variant to pick during primers designing?

[@StevenBradburn]

That depends on your research interest. Are you interested in a specific variant? Otherwise you can also design primers that amplify all variants

[@louisvandermerwe8012]

PCR is like Goldilocks, not too long not too short just right.

[@EcoGuna]

thank you :)

[@mirabella9043]

How can I design primers that cover several transcripts of one gene so I do not have to make a primer pair for each variant?

[@froylan026]

same question

[@khoopeikiet8863]

i intend to employ sybr green rt-pcr, so primer set that could "potentially"/not 100% complementarity binding to other unspecific region should be avoided? assume my gDNA is absence.

[@StevenBradburn]

Yes, that is correct :) try and avoid any off targets