NGS Setup Calculator

Search through our support documentation and FAQs for Archer, now part of Integrated DNA Technologies, products and services.

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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