Hello again Professor! With regards to the PCR, the machine says there is an amplification in my negative controls. But the CT values for the negative controls are consistently 10 points higher than the CT values for the actual samples. For example CT value of 37 for negative control vs CT of 21 for the sample. These results are still fine yeah?
I'm not really sure about your experimental context so can't really say by CT values alone. Have you run the samples on a gel to assess the size of the bands? Probably the next thing to try
Not always, depends on what you are trying to dilute out? Same principles apply when you are setting up your PCR mastermix and diluting it out for the different reactions.
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This is THE most thorough and effective PCR video i've ever seen
Thank you so much! Glad it helps
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The algorithm is tricky to figure out, but glad you were able to find the channel!
this is the clearest, most precise video I have seen on PCR. I normally do not comment but this was phenomenal
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Thank you! Took a long time to make, glad the effort shows on screen
Thank you for the simplified lesson. Keep up the good work.
Thanks for watching and commenting!
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Thanks for watching!
Absolutely Amazing
Thank you, this helped sooo much!!! You're videos are amazing!
Glad they help - thank you for taking the time to comment!
What amount of DNA should be loaded to each well?
Hello again Professor!
With regards to the PCR, the machine says there is an amplification in my negative controls. But the CT values for the negative controls are consistently 10 points higher than the CT values for the actual samples. For example CT value of 37 for negative control vs CT of 21 for the sample. These results are still fine yeah?
I'm not really sure about your experimental context so can't really say by CT values alone. Have you run the samples on a gel to assess the size of the bands? Probably the next thing to try
Do we need serial dilution for running a PCR?
Not always, depends on what you are trying to dilute out? Same principles apply when you are setting up your PCR mastermix and diluting it out for the different reactions.
OH THE COMB CREATES HOLES IN THE GEL
The gel lanes (or holes) need to be standard sizing all the way across - a comb makes that happen more or less