Hey, your videos amazing and very helpful, please can you make a video about how we do the analysis to the result after the pcr done by the thermocycler (biorad)...🙂
What’s the point of keeping the pcr plate on ice? Theres nothing in the mm that degrades at RT and most enzymes have hot start mechanisms to inhibit activity at RT
You would be right most of the time. Some enzymes out there function at lower temps like the NEAR enzyme. Also, there are thermodynamics to all reactions and colder temps limit activity even if it's small. It's a general good practice so you can maximize sensitivity and consistency of your assay.
@@Filtrous interesting! I did not know about NEAR PCR and it sounds interesting. There’s really 0 activity in hot-start enzymes mixes at RT but I get what you are trying to say
so what goes in the wells, how is the layout if we have standards and a reaction mix? Is it, reaction mix in all wells, and then in respective wells: cDNA of target genes, cDNA of housekeeping gene and cDNA of standards? i am so confused
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Future scientists NEED this! Tips and tricks are always appreciated.
Glad you appreciate this video David! What else would helpful for you?
JUST KEEP EVERYTHING ON ICE.
THAT WAS SO COOL.
Was quite a chilling experience.
Precise. Accurate. Perfect.
Thank you Navjeet, what other topic would be helpful for you that we could cover?
please can you make a video to setup full 384 well plate. how do you use Mult pipette for the run
Thanks... that was really very helpful... keep going man...✌
Thank you Yaya!
Hey, your videos amazing and very helpful, please can you make a video about how we do the analysis to the result after the pcr done by the thermocycler (biorad)...🙂
Great suggestion Yaya, I'll keep this in mind, thank you!
3000rpm... How much G is that on your centrifuge? Please remember that RPM are not standardised units (depends on centrifuge model), whereas G are.
Yes surr, not many people know this.
What’s the point of keeping the pcr plate on ice? Theres nothing in the mm that degrades at RT and most enzymes have hot start mechanisms to inhibit activity at RT
You would be right most of the time. Some enzymes out there function at lower temps like the NEAR enzyme. Also, there are thermodynamics to all reactions and colder temps limit activity even if it's small. It's a general good practice so you can maximize sensitivity and consistency of your assay.
@@Filtrous interesting! I did not know about NEAR PCR and it sounds interesting. There’s really 0 activity in hot-start enzymes mixes at RT but I get what you are trying to say
so what goes in the wells, how is the layout if we have standards and a reaction mix? Is it, reaction mix in all wells, and then in respective wells: cDNA of target genes, cDNA of housekeeping gene and cDNA of standards? i am so confused
or would you run housekeeping and target separately with their respective standards?