Documents & Support

Are your Nalgene containers tested for air shipment of liquid samples? Product FAQ

Answer

Most of our products are manufactured and tested for leakproof performance in laboratory and industrial settings only. Our PassPort IP2 bottles (2099 and 2199 series) represent the only exception to this: they are rigorously leak tested under a 15 psi (-95 kPa) pressure differential to simulate unpressurized air cargo transport.

Answer Id:: E15957

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My base spacing is too high/low. What does this mean and what can I do about it? Product FAQ

Answer

Base spacing is roughly defined as the number of scans per base. This value is directly related to the speed at which your samples are traveling through your polymer. The base spacing number will vary depending on the sample quality, capillary length, run module settings, polymer type, how long the polymer was on the system, electrophoresis buffer age, and environment. When you first start running samples, check the spacing to see what the average values are.

If the software is unable to determine the base spacing, it will give a best guess that will appear in red in the base spacing column. Usually this is either due to the data quality or a problem with the sample migration. If you notice the numbers fluctuating, check the following:

-Polymer expiration—if the polymer has expired, wash the block, syringe, and water/waste containers and replace.

-Polymer time on the system—the polymer should be in the syringe on the system for no more than a week. Polymer kept on the system longer than that may compromise the length of read and overall data quality.

-Electrophoresis buffer—the buffer is sold as a 10X concentrate and should be diluted to 1X prior to use. The 1X buffer is good for 1 month.

-Environment conditions—the instrument should be run at ambient temperature. Air flow can also affect migration. Make sure there are no fans, wind from an open window, or air vents blowing directly on the system.

-Sample quality—quantitate the template DNA prior to sequencing. After the sequencing reaction is done, clean the samples using one of the recommended procedures in the BigDye Terminator User Guide. If you are still getting poor results, run the Sequencing Standard as a sample to narrow the focus of your troubleshooting efforts. If the standard looks good, focus on the template and clean up procedures. If the standard looks bad, focus on the system.

If all of the above fail, please contact Thermo Fisher Scientific Technical Support for further assistance.

Answer Id:: E6256

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