The Features Of A Good qPCR Primer Pair
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- Опубліковано 2 сер 2024
- This video tutorial is part of the Mastering qPCR Online Course in The Top Tip Bio Academy.
🎓 MASTERING QPCR ONLINE COURSE 🎓
courses.toptipbio.com/p/maste...
In this video, I will share some of the features of a good PCR primer pair to take into consideration when designing primers. I have experience of performing hundreds of qPCR assays. In that time, I have gathered a lot of hints and tips. I will share my thoughts on good qPCR primer design.
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This is so wonderful! Save my time of struggling in grad school!
Thank you so much!!! :)
Can you make video about plasmid construction
thank you :)
Thanks for the video. If using gene-specific primers for quantification/detection, is it as necessary to bind towards the 3' end?
PCR is like Goldilocks, not too long not too short just right.
Actually, melting temperature is when 50% of a primer molecules is annealed to the target sequence, not just a half of it sequence!
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.
hi there, could you suggest which variant to pick during primers designing?
That depends on your research interest. Are you interested in a specific variant? Otherwise you can also design primers that amplify all variants
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.
Yes, that is correct :) try and avoid any off targets
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?
same question