Applied Biosystems™ Matrix Standards Kit, BigDye™ Terminator v3.1, for 31xx/3500/SeqStudio™/SeqStudio™ Flex

Catalog No.	Quantity
43-369-74	1 kit

Catalog No. 43-369-74 Supplier Applied Biosystems™ Supplier No. 4336974

Description

Includes

This kit contains two tubes of standard used to generate a spectrum required when analyzing BigDye Terminator v3.1 labeled DNA fragments on 31xx, 3500, SeqStudio, and SeqStudio Flex series genetic analyzers. Each tube contains a blend of four different colors of fluorescent dye-labeled DNA fragments suspended in 1X TE buffer.

This kit contains two tubes of standard used to generate a spectrum required when analyzing BigDye Terminator v3.1 labeled DNA fragments on 31xx, 3500, SeqStudio, and SeqStudio Flex series genetic analyzers.

This kit contains two tubes of standard used to generate a spectrum required when analyzing BigDye Terminator v3.1 labeled DNA fragments on 31xx, 3500, SeqStudio, and SeqStudio Flex series genetic analyzers. Each tube contains a blend of four different colors of fluorescent dye-labeled DNA fragments suspended in 1X TE buffer.

Order Info

1 Kit

Specifications

• Content And Storage: Store at 2–8°C protected from light. Do not freeze.
• Format: Kit
• For Use With (Equipment): 31xx Series Genetic Analyzers, SeqStudio™ Genetic Analyzer, 3500 Series Genetic Analyzers, SeqStudio Flex Series Genetic Analyzers
• For Use With (Application): Sequencing
• Label or Dye: ROX, TAMRA™, Rhodamine 110, Rhodamine 6G
• Product Line: BigDye Terminator, BigDye
• Product Type: Matrix Standard
• Quantity: 1 kit
• Shipping Condition: Wet Ice

Frequently Asked Questions (FAQs)

When I started my run on the Applied Biosystems 3500/3500xL Genetic Analyzer, an error occurred: "Unstable Electrophoresis current detected, check for air bubbles" but I don't see any bubbles. What else can cause this?

Some of the other causes of an Unstable Electrophoresis current detected error message are:

1.Leak on the system
2. Polymer that:
-Has expired
-Has been left on the instrument for more than the recommended time
-Is a mixture of expired polymer and non-expired polymer
3. Running Buffer that was:
-On the instrument longer than 2 weeks
-Not filled to the fill line or evaporated below the fill line
4. An arcing event that was not cleaned afterwards using the water wash wizard
5. Not performing regular maintenance on the instrument
6. Hardware issues

Inspect the system for leaks. If you do not see any leaks on the system, perform the wash the pump chamber and channels wizard using the conditioning pouch and place fresh, non-expired polymer and fresh Anode and Cathode buffer containers. If the problem persists, a service call may be required.

Do I need to replace the buffer every 14 days if the Applied Biosystems 3500/3500xL Genetic Analyzer is not in use?

If the instrument is not in use, it is not necessary to replace the buffer every 14 days. However, the buffer level needs to remain at the fill line to prevent the capillary or array from drying out. Top off the buffer volume with water if necessary. Replace the anode and cathode buffer containers prior to starting a new run.

How do I make a spectral standard for the Applied Biosystems 3130 series Genetic Analyzer?

The spectral calibration on the Applied Biosystems 3130 series systems can be performed by either using a Matrix Standard or a Sequencing Standard. The Matrix Standards will give 4 peaks, each of a different color whereas the Sequencing Standard is pre-sequenced and cleaned DNA that is dried down and just needs to be resuspended.

To make up the Matrix Standard:

1. Vortex the Matrix Standard tube for 1 min at full speed and pulse spin in a microcentrifuge.
2. Dilute the Matrix Standard in Hi-Di Formamide in a 1.5 mL microcentrifuge tube according to the following dilutions:
   For 4 capillary arrays
   
   • 36 cm and 80 cm arrays: mix 2.5 µL of Matrix Standard with 47.5 µL of Hi-Di Formamide.
   • 50 cm array: mix 2.5 µL of Matrix Standard with 97.5 µL of Hi-Di Formamide.
   
   
   For 16 capillary arrays
   
   • 36 cm and 80 cm arrays: mix 10 µL of Matrix Standard with 190 µL of Hi-Di Formamide.
   • 50 cm array: mix 5 µL of Matrix Standard with 195 µL of Hi-Di Formamide.
3. Mix thoroughly and pulse spin in a microcentrifuge.
4. Denature at 95 degrees C for 2 min and dispense 10 µL of reagent into wells A1–H12 (16 capillaries) or A1–D1 for 4 capillaries on a 96-well plate.
5. Briefly spin to ensure there are no air bubbles at the bottom of the well.
6. Set up and run the spectral calibration as per the Applied Biosystems 3130/3130xl Genetic Analyzers Getting Started Guide

More information on making up the Matrix Standards can be found in the BigDye Terminator v.3.1 Matrix Standard Kit for 3130 and 3100 Series Systems Product Insert or the BigDye Terminator v.1.1 Matrix Standard (Dye Set E) for 3100, 3130, and 3500 Series Genetic Analyzers Product Insert

To make up the Sequencing Standard:

1. Resuspend one tube of the Sequencing Standard with 170 µl of Hi-Di Formamide.
2. Vortex at full speed for 1 min and pulse spin in a microcentrifuge.
3. Denature at 95 degrees C for 2 min and dispense 10 µL of reagent into wells A1–H12 (16 capillaries) or A1–D1 for 4 capillaries on a 96-well plate.
4. Briefly spin to ensure there are no air bubbles at the bottom of the well.
5. Set up and run the spectral calibration as per the Applied Biosystems 3130/3130xl Genetic Analyzers Getting Started Guide

For Research Use Only. Not for use in diagnostic procedures.