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  • illumina primer/adaptor concentrations

    Hi all,

    I want to switch from illumina supplied library prep kit to assembling my own. I've been seeing helpful threads about primer/adapter sequences, but cannot find any information about their concentrations.

    I am specifically interested in concentration of SE lib prep kits:
    PCR primers 1.1/2.1
    Adapter mix

    Does anyone know these concentrations provided in the kit (or point me to threads containing these information)?

    Thanks!
    Last edited by tamosan; 10-19-2009, 04:17 PM.

  • #2
    Oligo concentrations

    Originally posted by tamosan View Post
    I am specifically interested in concentration of SE lib prep kits:
    PCR primers 1.1/2.1
    Adapter mix

    Does anyone know these concentrations provided in the kit (or point me to threads containing these information)?

    Thanks!
    The concentration in the adapter mix is 15 uM - you can find this info in the Cronn et al 2008 paper in NAR, although you can also work it out from the Illumina protocol: for DNA fragmented to an average size of ~500bp, 5 ug in 10ul gives a total of 15 picomoles; therefore for a 10:1 molar ratio, you must be putting in 150 picomoles of adapter (also in 10ul).

    For the PCR primer concentration, the thread "Excess PCR primers in final product " has posts indicating the stock PCR primers to be 25uM; these are used 1 in 50 in the enrichment PCR, i.e. at a final concentration of 500nM.

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