Illumina library quantification using a QuantStudio 5

[tatresi]

Hello,

Does anyone have any successful experience using a QuantStudio 5 to quantify their illumina libraries? I've been trying to get decent results using the KAPA Rox low qPCR kit as recommended by KAPA. The standard curve looks fine but I'm getting much fewer reads than I would after quantification on our 7900.

[SequaPodPerson]

Can you clarify a couple things? You've used the KAPA kit with success on the 7900 or you were using a different kit/homebrew?

When you say reads, do you mean sequencing reads after loading based on the KAPA data or you mean the KAPA is giving you lower-than-expected concentrations?

Have you tried making one master plate and stamped it out into two plates to do a head-to-head between machines?

I'm just at the end of a couple of long weeks of troubleshooting this kit and in the end, it was the machine (a ViiA 7) so it's definitely a possibility!