cDNA synthesis and statistical variation - (Aug/10/2007 )
Hello,
I needed to prepare cDNA from more than 50 samples. However, it was not possible to fit all my samples in one block of the PCR machine in the lab. Thus, i decided to prepare all my samples at the same time and just split the samples in two blocks and run the same programe at the same time.
Now that i am thinking of it again i realized that i run the control group (reference group) only in one of the blocks. Although i think that the machine will not give so much of a variation i think that the right way to do it would be to prepare again the cDNA by running the control twice, on the two different blocks.
Another PhD student in the lab told me that in a systemiatic basis she runs the control only in one of the blocks and then she compares all her groups with that even the control was not in the block during the runm, but all the samples they have been prepared and run at the same time. Could she be right?
What is your opinion on that?
Thank you in advance for the feedaback,
I needed to prepare cDNA from more than 50 samples. However, it was not possible to fit all my samples in one block of the PCR machine in the lab. Thus, i decided to prepare all my samples at the same time and just split the samples in two blocks and run the same programe at the same time.
Now that i am thinking of it again i realized that i run the control group (reference group) only in one of the blocks. Although i think that the machine will not give so much of a variation i think that the right way to do it would be to prepare again the cDNA by running the control twice, on the two different blocks.
Another PhD student in the lab told me that in a systemiatic basis she runs the control only in one of the blocks and then she compares all her groups with that even the control was not in the block during the runm, but all the samples they have been prepared and run at the same time. Could she be right?
What is your opinion on that?
Thank you in advance for the feedaback,
It is always better to include the controls. It may be completely unecessary, but it never hurts (except in terms of cost) and it might uncover a problem that you had no idea existed.