NuQuant Assay for Qubit:

The NuQuant library quantification method accurately measures molar concentrations of NGS libraries. NuQuant library quantification is a proprietary method whereby a specific number of fluorescent labels are incorporated into the library molecules during library preparation. Consequently, each library molecule has an equivalent number of labels incorporated, regardless of the size of the library fragment. The molar concentration of sequenceable molecules can be directly measured using fluorometers such as Qubit® or standard plate readers.

Downloading the application

Choose the appropriate instrument file below for the NuQuant product you are using.

Installation instructions are provided below.

To download files to your local disk, right-click the download link and select “Save as”.

NuQuant for DNA-Seq kits

Kit	App name	Qubit 2.0	Qubit 3.0 or 4
Celero DNA-Seq Mech Library Preparation	NuQuant 644	download	download
Celero DNA-Seq Enz Library Preparation	NuQuant 644	download	download
Celero DNA-Seq (legacy kit)	Celero PCR	download	download
Celero EZ DNA-Seq (legacy kit)	Celero PCR	download	download

NuQuant for RNA-Seq kits

Kit	App name	Qubit 2.0	Qubit 3.0 or 4
Revelo RNA-Seq High Sensitivity	NuQuant 644	download	download
Universal Plus mRNA-Seq	Univ. Plus	download	download
Universal Plus Total RNA-Seq	Univ. Plus	download	download
Universal RNA-Seq (legacy kit)	Univ. Plus	download	download

Installing the application

Installing NuQuant on Qubit 2.0

Note: NuQuant can only be implemented on Qubit® 2.0 Fluorometers running firmware v3.11 (or higher). For instruments requiring an upgrade, see firmware upgrade instructions here.

1. After downloading the file Qubit2<KIT NAME>v1.qbt from the above link, copy it to the root directory of a USB drive (do NOT place the .qbt file into a folder in the USB drive).
   • Important: Make sure there is only one .qbt file saved to the root directory of the USB drive. Having multiple .qbt files present on the USB drive will result in an error when uploading the file.
2. Unplug the Qubit power source, insert the USB drive containing the Qubit2<KIT NAME>v1.qbt file into the USB drive port of the Qubit® 2.0 Fluorometer, and plug the power back in.
3. You will see the message *“Qubit2v1.qbt file detected. Do you wish to upload?"* Select `Yes`.
4. On the home screen you will now see a button marked <KIT NAME>. Select this to perform the NuQuant assay for NuQuant library products.

Installing NuQuant on Qubit 3.0 and Qubit 4

1. After downloading the file Qubit3and4<KIT NAME>.qbt from the above link, copy it to the root directory of a USB drive (do NOT place the .qbt file into a folder in the USB drive).
   • Important: Make sure there is only one .qbt file saved to the root directory of the USB drive. Having multiple .qbt files present on the USB drive will result in an error when uploading the file.
2. Insert the USB drive into the USB drive port of the Qubit® 3.0 or Qubit® 4 Fluorometer.
3. Go to the Settings screen and touch Import new assay. The instrument searches the USB drive for the .qbt file and the Import new assay screen displays “ NuQuant " under Assay name.
4. Touch to “ NuQuant ” to select. It will become highlighted. Touch `Next`.

   • Option A: Select a destination folder for the new assay and touch Save. Each folder can hold up to 9 assays.

   • Option B: Alternatively, touch New folder to create a new destination folder. On the New folder screen, touch the New folder text field. A keyboard pops up. Using the keyboard, type in the name of the new destination folder. You can use up to 11 characters; spaces at the beginning or the end of the name are not recognized. Touch Enter, and then touch Done to return to the “Import new assay – Where” screen. The newly created folder will appear in the list of folders. Select the new destination folder and touch Save.

5. On the verification screen, touch Done to complete the import process.
6. On the home screen you will now see a button marked . Select this to perform the NuQuant assay for NuQuant library products.

Uninstalling NuQuant on Qubit 2.0

If you wish to remove NuQuant, including the <KIT NAME> button from your Qubit home screen, perform the following steps:

1. Follow the steps for upgrading firmware (see firmware upgrade instructions.)
2. Once you press the Update button, a message will appear titled “Updating: Select Assay(s) to Delete”. Select <KIT NAME>. The ** line will turn gray, indicating it has been selected.
3. Click OK.
4. Another window will appear with the message “The selected assay(s) will be deleted. Do you wish to proceed?” Click Yes.
5. The “Updating firmware…” status bar will appear. After approximately 1 minute, the update will be complete and the Tecan NuQuant app will have been removed.

Uninstalling NuQuant from Qubit 3.0 and Qubit 4

1. Go to the Settings screen and touch Manage assays.
2. Touch Show/hide Assays.
3. Select the folder which contains NuQuant.
4. Next to <KIT NAME> NuQuant <version>, touch Delete, then Done.
5. The next screen will ask “This will permanently delete the selected assay(s). Do you wish to continue?” Touch OK.