Am I required to use fresh PCR product with the BigDye Direct Cycle Sequencing Kit, or can I store the PCR reaction before proceeding? What temperature is best for storage?
We always recommended that you use fresh PCR product, but if you need to store it, the amplified PCR reaction can be stored for 16 hours at 4°C or up to 4 weeks at –20°C.
Can I store samples in BigDye Xterminator solution? If so, what is the optimal temperature, and for how long can I store it?
Yes, after cleaning the sequencing product using the BigDye XTerminator Purification Kit, the end product can be stored up to 10 days at either 4°C or –20°C.
If I need to pause my experiment in the middle of the BigDye Direct Cycle Sequencing Kit procedure, how long can I expect my samples to be stable?
The amplified PCR reaction can be stored for 16 hours at 4°C or up to 4 weeks at –20°C. The formulated sequencing reaction mix can be stored at ambient temperature for up to 16 hours. After cycle sequencing but before cleanup, the sequencing product can either be stored for up to 16 hours at 4°C or up to 4 weeks at –20°C. After cleaning the sequencing product up using the BigDye XTerminator Purification Kit, the end product can be stored up to 10 days at either 4°C or –20°C.