# Normalization Buffer Volumes

**Available from:** Clarity LIMS v1.0

This section discusses the *normalizationBufferVolumes* script, which is used to calculate buffer volumes.

## Script Overview

The *normalizationBufferVolumes* script generates a comma-separated file — Normalization buffer volumes.csv — file that holds the calculated buffer volumes information. The generated file is a attached to the Library Normalization step in the LIMS.

The script calculates the 'Volume of Dilution Buffer (uL)' as follows:

* "Volume of Dilution Buffer (uL)" = ( ("Source Volume (uL)" \* "Source Conc.") / "Destination Conc." ) - "Source Volume (uL)"

{% hint style="danger" %}
The script

* requires that input samples to the step have Concentration and Conc. Units UDFs.
* only supports Conc. Units of nM.
  {% endhint %}

## Script Parameters

| **Parameter** | **Description**                                             |
| ------------- | ----------------------------------------------------------- |
| -u {username} | (Required) LIMS login username (Requires admin credentials) |
| -p {password} | (Required) LIMS login password                              |
| -i {URI}      | (Required) LIMS process URI                                 |
| -c {filename} | (Required) Normalization Buffer Volumes CSV file name       |

## Command Line Example

<details>

<summary>Example</summary>

{% code overflow="wrap" %}

```markup
bash -l -c "/opt/gls/clarity/bin/java -jar \
/opt/gls/clarity/extensions/ngs-common/v5/EPP/ngs-extensions.jar script:normalizationBufferVolumes \
-i {processURI:v2:http} \
-u {username} \
-p {password} \
-c {compoundOutputFileLuid0}
```

{% endcode %}

</details>

## Configuration

The following step UDFs are configured on the Library Normalization step:

* Library volume (ul) transferred to DESTINATION plate: Numeric, Default = 10
* Normalized conc. (nM): Numeric, Default = 2
* Maximum destination container volume (ul): Numeric, Default = 800

The inputs to the step must have values for the following sample UDFs:

* Concentration: Numeric
* Conc. Units: Single Line Text

## Rules & constraints

* One sample input generates one sample output and one shared result file.
* The CSV file header contains the date and a record of the user who generated the file.
* The contents of the CSV file are ordered by source plate and then source well.

## Additional information

Other scripts you may find useful:

* [Initialize Artifact UDFs](https://help.claritylims.illumina.com/instruments-and-integrations/ngs/ngs-initialize-artifact-udfs)