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# How to prepare low concentration (< 2 nM) libraries for sequencing on a MiSeq
The [MiSeq Illumina Denature and Dilute Protocol Generator](https://support-docs.illumina.com/IN/dnd-wizard/Content/dnd/landing.htm?ins=miseq) has directions for denaturing 2 nM and 4 nM libraries with the [MiSeq Standard Normalization Method](https://support-docs.illumina.com/IN/MiSeq_DnD/Content/MiSeq/DnD-MiSeq.htm?protocol=standard).
* For denaturation steps, make sure that the concentration of NaOH is no more than 1 mM in the final solution diluted with HT1. Higher NaOH concentrations can inhibit library hybridization to the flow cell, leading to decreased cluster density.
Libraries that are less than 2 nM but at least 0.5 nM can be prepared for MiSeq sequencing. This is done by following the Standard Normalization Method in the [Denature and Dilute Libraries for the NextSeq 500 and NextSeq 550 Sequencing Systems](https://support-docs.illumina.com/IN/NextSeq550_DnD/Content/NextSeq550/DnD-NS550.htm?protocol=standard) Protocol Generator until libraries are diluted to 20 pM. In this protocol, excess NaOH is offset by adding 200 mM Tris-HCl, pH 7 to make sure that NaOH is fully hydrolyzed in the final solution.
* To denature < 2 nM libraries, follow the steps for starting library concentration of 1 nM or 0.5 nM in the [Denature and Dilute Libraries for the NextSeq 500 and NextSeq 550 Sequencing Systems](https://support-docs.illumina.com/IN/NextSeq550_DnD/Content/NextSeq550/DnD-NS550.htm?protocol=standard), see **Figure 1**. When libraries are at 20 pM, proceed to the Dilute Denatured 20 pM Library step in the [Denature and Dilute Libraries for the MiSeq system](https://support-docs.illumina.com/IN/MiSeq_DnD/Content/MiSeq/DnD-MiSeq.htm?protocol=standard) under the section "Denature and Dilute, 4 nM Libraries", see **Figure 2**.
**Figure 1**.Denaturation and Dilution steps for Standard Normalization Method in the NextSeq 500/550 denature and dilution workflow.
**Figure 2**.Dilute Denatured 20 pM Library step for the Standard Normalization Method in the [Denature and Dilute Libraries for the MiSeq system](https://support-docs.illumina.com/IN/MiSeq_DnD/Content/MiSeq/DnD-MiSeq.htm?protocol=standard). To add a PhiX control, proceed to the "Denature and Dilute PhiX Control (Optional)" section of the [Denature and Dilute Libraries for the MiSeq system](https://support-docs.illumina.com/IN/MiSeq_DnD/Content/MiSeq/DnD-MiSeq.htm?protocol=standard).
Refer to the specific [Denature and Dilute Protocol Generator](https://support.illumina.com/downloads/denature-and-dilute-protocol-generator.html) for specific instruments for detailed instructions.
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