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.

Select platform:

Library pool concentration (nM):

Desired library pool concentration (nM):

NextSeq/MiSeq loading pool

Desired loading concentration (pM)

Desired loading volume (μL)	1000

Desired PhiX fraction (%):

PhiX stock concentration (pM):

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:

HT1 Buffer:

PhiX:

Cartridge:

Flow cell:

Incorporation buffer:

Print for your records

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