Editing BioMicroCenter:RTPCR Protocol for Sample QC (section)

== Using RT-PCR Information to Improve Clustering ==

We utilize the concentrations obtained by RT-PCR to help better control the amount of sample that we are loading onto each flowcell for sequencing. We take these concentrations into account during the denaturation step of cluster generation.  2uL of a 10nM sample will be denatured in 6uL EB and 2uL of 0.5N NaOH for a final volume of 10uL.  

For samples that are under 10nM:

* Adjust sample:EB ratio to allow for proper loading concentration onto each flowcell. If samples are not concentrated enough, you may boil your samples at 98°C for 5min as an alternative to the sodium hydroxide denaturation. 

For samples over 20nM:

* Dilute samples down to 10nM with EB and follow NaOH denaturation recipe above.