3 replies to this topic

[thekid]

Hello,

Is there any way to analyze real time SYBR green assays done on 2 different machines (ABI 7500 and Bio-Rad). Plasmid control with similar copy numbers were used in both machines. The machine calculated threshold was taken for all calculations.

    Can the results be merged and analyzed or will there be difference since the algorithm used by the machines differ in its calculations?

Or does the use of standard plasmid copy number in both machines negate the problem with software analysis?


Thank you for the replies,
thekid.

[Curtis]

data will be different. I have done this before.

[thekid]

Hi,

  thank you for the response. Have you come across any papers that presents data from two different machines?

thekid

[biznatch]

It depends what's done on each machine.  For example let's say you have two genes (X and Y) and two treatments (C and KO) and you want to know how the expression of each gene is changed in C vs KO.  I would make sure that both C and KO from gene X are always run together (same machine and at the same time), but you could run C and KO from gene Y on a different machine or on the same machine but at a different time.  If gene X is increased 3 fold in KO over C, it shouldn't really matter what machine you use, it should always be 3 fold increased.  Though as with anything there may be slight variations.  I definately wouldn't run C on one machine and KO on the other and try to compare them.

Edited by biznatch, 28 May 2012 - 05:52 PM.