I'm having to give an R coding walk-through to some Stanford first years on Wednesday, and it had been so long since I'd done any qPCR I had forgotten the details. This video was perfect. Thanks for making such a well organized, clear and informative video.
eventually however enough amplified product accumulates to generate a 1:22detectable signal the cycle number of which this occurs is the threshold cycle 1:27or CT in this plot the threshold fluorescence is indicated by the green 1:32line in real-time quantitative pcr the data that are generated are the 1:42threshold cycle or CT values the threshold cycle is determined mainly by 1:48the amount of template present at the start of the amplification reaction if a 1:52large amount of template is present at the start of the reaction relatively few
Great video! I went through 5 other videos that were either too simple, way too drawn out to keep watching, or ended up being sales pitches for certain reagents or instruments.
Hello. Can anybody help me to calculate how much fold increase/decrease in gene of my interest occurred as a result of my compound treatment. I have been trying to sort it out but couldn't . Ct values for GADPH and WNT of control group are 14.3292 and 22.274 respectively, while Ct values for GAPDH and WNT in treated group are 14.1418 and 23.9879 respectively. Please help me to reach to the conclusion. Regards!
So in absolute quantification, you determine how many copies of the sequence are in your sample, but how would you translate that information into how many copies are in the genome?
Shouldn't we take the ratio as calibrator/test instead of test/calibrator in order to conclude that expression of the gene is 8-fold higher in tumor cells, since the ratio of 8 indicates that "test"/tumor cells have higher CT, thus lower expression? I'm just checking because I'm new to the subject, I'd appreciate it if someone could answer :)
if we want to know significant value from expression gene by statistic. What should i have to input in spss? Value from 2-∆∆Ct from sampel treatment and control? Or value from ∆CT sample and control
At the present, My Piko Real PCR system (Thermo Scientific) had E36 error (Strain gauge has problem). Could you help us to solve the error? Hope to get your feedback.
when i go for absolute quantification of DNA, after the experiments green coloured flags are apperas on it, along with standard samples and with unknown samples, may i know why this happens sooo ? i am using ABI Stepone plus ver2.2. kindly help or suggestions r welcome
may i know what is the unit of quantity while quantification of pathogen load ? another how to convert teh absolute data into to copy no. of the pahogen.? kinldy help urgently
Why is it truncated this video? what about the normalization against the reference gene? Since is actually the video of another video (BioRad's ?) you could have referentiated it, right? Disappointing...
Hi Asif-I have forwarded your request to Bio-Rad Tech support. Hopefully I will have an answer for you shortly. You can also write to them directly by filling out the email support form on this page www.bio-rad.com/en-us/life-science-research/support
I'm having to give an R coding walk-through to some Stanford first years on Wednesday, and it had been so long since I'd done any qPCR I had forgotten the details. This video was perfect. Thanks for making such a well organized, clear and informative video.
eventually however enough amplified product accumulates to generate a
1:22detectable signal the cycle number of which this occurs is the threshold cycle
1:27or CT in this plot the threshold fluorescence is indicated by the green
1:32line in real-time quantitative pcr the data that are generated are the
1:42threshold cycle or CT values the threshold cycle is determined mainly by
1:48the amount of template present at the start of the amplification reaction if a
1:52large amount of template is present at the start of the reaction relatively few
What do you do when the control Ct is undetermined, like in a case of transgenic line versus WT where a foreign gene is undetected in the WT.
I didn't take any control gene,how to calculate delta delta value now? suggestions plz
Great video! I went through 5 other videos that were either too simple, way too drawn out to keep watching, or ended up being sales pitches for certain reagents or instruments.
What is the app which help to take a video?
Your content is so touching
Hello. Can anybody help me to calculate how much fold increase/decrease in gene of my interest occurred as a result of my compound treatment. I have been trying to sort it out but couldn't .
Ct values for GADPH and WNT of control group are 14.3292 and 22.274 respectively, while Ct values for GAPDH and WNT in treated group are 14.1418 and 23.9879 respectively.
Please help me to reach to the conclusion.
Regards!
So in absolute quantification, you determine how many copies of the sequence are in your sample, but how would you translate that information into how many copies are in the genome?
how did you do it can you share with me , thank you
Shouldn't we take the ratio as calibrator/test instead of test/calibrator in order to conclude that expression of the gene is 8-fold higher in tumor cells, since the ratio of 8 indicates that "test"/tumor cells have higher CT, thus lower expression? I'm just checking because I'm new to the subject, I'd appreciate it if someone could answer :)
You are right!! Had the same question and I think they are missing a minus sign in the formula given at 9:30
if we want to know significant value from expression gene by statistic. What should i have to input in spss? Value from 2-∆∆Ct from sampel treatment and control? Or value from ∆CT sample and control
At the present, My Piko Real PCR system (Thermo Scientific) had E36 error (Strain gauge has problem). Could you help us to solve the error? Hope to get your feedback.
I ve got some results with + not in mince what does that mean
when i go for absolute quantification of DNA, after the experiments green coloured flags are apperas on it, along with standard samples and with unknown samples, may i know why this happens sooo ? i am using ABI Stepone plus ver2.2. kindly help or suggestions r welcome
can i use this relative quantification method in telomere length by using ratio
How to calculate log value.
may i know what is the unit of quantity while quantification of pathogen load ? another how to convert teh absolute data into to copy no. of the pahogen.? kinldy help urgently
Why is it truncated this video? what about the normalization against the reference gene?
Since is actually the video of another video (BioRad's ?) you could have referentiated it, right?
Disappointing...
There's a part2 of this video...
HBV DNA detected within the liner range of the assay means
Great explanation. Thank you very much!
Hello! Can I help you translate this video to Spanish?
How can i know the efficiency is 100%.
you don't. it is an assumption.
Please help Bio-Rad
Hi Asif-I have forwarded your request to Bio-Rad Tech support. Hopefully I will have an answer for you shortly. You can also write to them directly by filling out the email support form on this page www.bio-rad.com/en-us/life-science-research/support
americanbiotech Thanks for prompt response. I will definitely fill out the form as well as keep on visiting this page for the reply from Tech support.
very helpful thanks
interpolate, not extrapolate!
Thank you
very clear
R.T PCR with milk samples
Thompson Robert Wilson Jose White Deborah
thank you very much
Nice! Thank you :)
Dat voice
not helpful at all
You are exerciseng torture on living men and women this breaking every law and legally any were in world or iso or any digit sumbal place of existence