NGS Setup Calculator

The easy way to calculate how much of your Archer library to load on a sequencer

This tool assists in the creation of a library pool ready for sequencing on an Illumina® MiSeq® or Illumina NextSeq® instrument.

*To use this calculator, pool final library concentrations between 5 and 20nM and quantify using the KAPA Universal Library Quantification Kit from KAPA Biosystems.

NextSeq/MiSeq loading pool
Desired loading volume (μL) 1000

1) Dilution of library pool to 4nM
Library pool volume (µL): 8.0
Add H2O (µL): 2.0
Library Conc. (nM) NaOH Conc. (mM)
4 0.0


2) Denature library pool
Library pool from Step 1 (µL): 10.0
200mM NaOH (µL): 10.0
Incubate 5min at room temperature
Library Conc. (pM) NaOH Conc. (mM)
2000 100.0


3) Neutralize and dilute library pool
Denatured input from Step 2 (µL): 20.0
200mM Tris, pH 7.0 (µL): 10
HT1 Buffer (µL): 970.0
Library Conc. (pM) NaOH Conc. (mM)
40 2.0


4) Create aliquot to load onto MiSeq/NextSeq cartridge
Denatured pool from Step 3 (µL): 375.0
PhiX (µL) 78.9
HT1 Buffer (µL): 546.1
Final Pool (pM) Final NaOH (mM)
15 0.750


5) Load final pool into cartridge

For record-keeping purposes, record reference/lot numbers and expiration dates here:

Print for your records

For Research Use Only. Not for use in diagnostic procedures.  For Research Use Only. Not for use in diagnostic procedures.

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