Protocol Online logo
Top : New Forum Archives (2009-): : PCR, RT-PCR and Real-Time PCR

iCycler Calibration Question - (Jan/24/2013 )

Hi, I need to calibrate an iCycler rt-pcr machine for SYBR green I use. The documentation is very vague for this and the iCycler calibration kit I have does not include a SYBR green standard. What do I use for the standard? is a standard (pure dye) even needed? The SYBR green that I have also has fluorescein in the mix.

If anyone uses a bio-rad iCycler or IQ5 with SYBR green and knows how to calibrate it, please let me know!



Bio-Rad did get back to me. To calibrate an iCycler for SYBR green, all you need to do is copy the .RME file for FAM and paste in the value for SYBR-490 in the .RME file in the program folder. You can do this since FAM and SYBR green use the same wavelengths.

It is not intuitive at all and they definitely don't mention this in the users manual.