MyQubit—
Envision and Create New Assays For the Qubit® 2.0 Fluorometer

MyQubit brings your favorite fluorescence assays right to your benchtop, providing a reliable platform for many quantitation needs—from laboratory research and quality control to process monitoring and beyond. Any reagent or assay that is spectrally compatible with the Qubit® hardware can be adapted for use with the Qubit® 2.0 Fluorometer.

Download MyQubit firmware update & instructions

Learn more about the Qubit® Fluorometer

About the MyQubit Firmware Update

Create new Qubit® applications

MyQubit allows you to create new applications for the Qubit® 2.0 Fluorometer in minutes using parameters that can easily be uploaded to the instrument using a USB drive, without changing the existing assays.  All new Qubit® 2.0 instruments are now pre-loaded with the new MyQubit firmware and current owners can download the firmware.


Easy setup

Since the instrument is operated by simple components, creation of additional applications is as straightforward as matching spectral compatibility with the LEDs and emission filters used.  The instrument is operated by two LEDs: blue with a maximum around 470 nm, and red with a maximum around 635 nm (filters 430 – 495 nm and 600 – 645 nm, respectively).  Two emission filters are set from 510 – 580 nm and 665 – 725 nm.  New assays can be based on existing Life Technologies assays, modifications/optimization of existing reagents, or completely novel ideas altogether.

 

How to Create Your Own Assay Using MyQubit

Overview

Each MyQubit file can store information for up to four unique assays.  The MyQubit file template* includes parameters for a single assay.  To add additional assays, simply copy and paste the appropriate fields below the entries for assay “Name 1.”  Additional assay name headers must be defined as “Name 2”, “Name 3” and “Name 4.”  Although all fields may not be populated for every assay, all field headers must be included for each assay in the MyQubit file in order for the file to upload properly.  The "MyQubit File Parameters & Definitions" table below describes the different fields that comprise a MyQubit file and provides guidelines for defining them.

Step 1: Enable Raw Mode on Your Qubit® 2.0 Fluorometer

The Qubit® 2.0 Fluorometer can be now used as a mini-fluorometer with Qubit® Raw mode (right click, choose 'Save As').

First save the Qubit_Raw.qbt file to your PC desktop and then copy to your USB drive.  To upload Qubit® Raw mode, first turn the power off by unplugging the Qubit® 2.0 Fluorometer.  While the instrument is unplugged, insert the USB drive into the instrument and plug back in.  Your Qubit® 2.0 Fluorometer will recognize if the Qubit_Raw.qbt file is present on the USB drive and you will be prompted to upload the).  Once upload is complete, you will be directed to a new Home Screen where the RAW function is now accessible.

Step 2: Collect Data and Define Constants for MyQubit Assays

Collecting the data

Qubit® Raw mode allows you to manually select the excitation light source of your choice, i.e. blue LED, red LED, both or none, while reading fluorescence in both the green and far red emission channels.  Raw fluorescence values are obtained to generate data for the development of new assays, or to validate the performance of existing ones. We recommend, as starting point formatting assays as defined in the “MyQubit EZ Guide” spreadsheet.

Defining the constants

In addition to the straightforward parameters such as assay name and desired units output, several key constants must also be resolved prior to creating a new assay.  These constants define the shape of the standard curve used in calibration.   For your convenience, we have developed the “MyQubit EZ Guide” Excel® spreadsheet that can be used to generate these constants.

Step 3: Define your assay

Creating the MyQubit file
To create your own MyQubit file, use a simple word processor such as Notepad (Figure 1).  The file must be saved with the extension .qbt.  MyQubit files should first be created and saved external to the Qubit® 2.0 Fluorometer on a PC and then transferred to the USB drive provided with the instrument.  The name of the file should not contain more than 20 characters from the standard ASCII library, excluding commas.  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 to the Qubit®2.0 Fluorometer.

Download the MyQubit .qbt file template*

*MyQubit file download instructions: 1) Click to download link; 2)  save to PC;  3)  Open with a simple text editor such as Notepad, edit and re-save;  4)  Upload to Qubit® with USB.
 

Figure 1. Example of a .qbt file.

Step 4: Upload your assay

To upload your new assay, first turn the power off by unplugging the Qubit® 2.0 Fluorometer.  While the instrument is unplugged, insert the USB drive into the instrument and plug back in.  Your Qubit® 2.0 Fluorometer will recognize if there is a .qbt file present on the USB drive.  If the file is correctly populated, you will be prompted to upload the assays contained in the .qbt file (Figure 2).  Once upload is complete, you will be directed to a new Home Screen where the Parent Name of your new MyQubit assay is visible.



Figure 2.  MyQubit will guide you through the process of uploading new assays.  Up to 20 new assays can be permanently uploaded without affecting the look or performance of existing applications.


Step 5: Perform your assay

Your new MyQubit assay is ready to use.  Follow the guidelines for existing assays to begin use. Please refer to the Qubit® 2.0 Fluorometer User Manual for a detailed description of instrument function and assay workflow.

MyQubit File Parameters & Definitions

Parameter Accepted Values Description
Parent Name(user defined, max 10 characters)Indicates the name visible on the initial Home Screen, i.e. the “family” of assays.  For example, “DNA” is a Parent Name for the existing instrument.
Name1 (and Name2, 3, 4)(user defined, max 10 characters)Indicates the individual assay name.  For example, “dsDNA” for the existing instrument.
Name Sub-text(user defined, max 20 characters)Indicates any smaller sub-text under the assay name (or to the right of the assay name from the graphical display).  For example, “High Sensitivity” for the existing instrument.
CalibrationYES or NOIndicates whether you are creating an assay that requires calibration or not.  “YES” indicates that the assay will require calibration.  All assays created using MyQubit that require calibration will utilize two standards, the first of which is always a sample blank (analyte concentration of “0”) and the second of which is a sample of a concentration to be determined by the user. “NO” indicates that the assay will not require calibration.  In this case, the output will be a raw fluorescence value or values. ACCEPTED VALUES
Units(user defined, text only, max 10 characters)Only required when Calibration is “YES.”  Maximum length 10 characters.
ExcitationBLUE, RED, BOTH, or NONEBOTH and NONE are only acceptable if Calibration is “NO.”  Otherwise, this constitutes an error and the .qbt file will not upload. BLUE = 470 nm, filter 430 – 495 nm. RED   = 635 nm, filter 600 – 645 nm. ACCEPTED VALUES
EmissionGREEN, FAR RED, or BOTHBOTH is only acceptable if Calibration is “NO.”  Otherwise, this constitutes an error and the .qbt file will not upload. GREEN = 510 – 580 nm. FAR RED = 665 – 725 nm. ACCEPTED VALUES
Std Signal Ratio Limit(user defined, numeric only)Only needed when Calibration is “YES.” This number specifies the amount, in raw fluorescence, that the signal for Standard #2 must exceed the signal for Standard #1 for your MyQubit assay.  This is meant to help troubleshoot large oversights in calibration.  If you are unsure of a reasonable threshold, or if this is not of concern to you, then enter a small value that you will be assured of attaining in most calibrations.
Low(user defined, numeric only)Only needed when Calibration is “YES.”  Indicates the lowest sample concentration that the instrument will display a result for.  Samples with concentrations below this value will be reported as “< Low” and will display the message “*Sample TOO LOW*”
High(user defined, numeric only)Only needed when Calibration is “YES.”  Indicates the highest sample concentration that the instrument will display a result for.  Samples with concentrations above this value will be reported as “> High” and will display the message “*Sample TOO HIGH*” 
N(user defined, numeric only)Only needed when Calibration is “YES.”  One of three algorithm constants that must be determined by the user.  Indicates the linearity of the curve, with a value of 1.00 corresponding to completely linear.
K(user defined, a scaling factor, numeric only)Only needed when Calibration is “YES.”  One of three algorithm constants that must be determined by the user.  Indicates the portion of the curve to focus on. 
S(user defined, numeric only)Only needed when Calibration is “YES.”  One of three algorithm constants that must be determined by the user.  Indicates the concentration value of Standard #2 used in assay calibration.
Green RFU Name(user defined, max 20 characters)Indicates the name given to the displayed RFU output for Green emission when Calibration is “NO.”  Only needed when Calibration is “NO” and Emission is “GREEN” or “BOTH.”
Far Red RFU Name(user defined, max 20 characters)Indicates the name given to the displayed RFU output for Far Red emission when Calibration is “NO.”  Only needed when Calibration is “NO” and Emission is “FAR RED” or “BOTH.”