Description:
The BigDye^® Terminator 5X Sequencing Buffer reduces sequencing costs without affecting sequence accuracy or read length. Its buffer system enables to use less dye terminator mix.

Applications:

• DNA sequencing
• Fragment analysis

Recommended Storage Condition: 4°C

Recommended Protocol:
Prepare sequencing reactions (10µl rxn) according to the following:
- Add dye terminator mix 0.25-0.5µl
- Add 5X Sequencing Buffer 2µl
- Add template (100-200 ng/µl) 1 µl
- Add primer (3 pmol/µl) 1 µl
- Add H₂O to 10 µl
Cycle the reaction according to the following protocol:

Step 1: 96°C  3 minutes
Step 2: 96°C  10 seconds
Step 3: 50°C  5 seconds
Step 4: 60°C  2 minutes and 30 seconds
(25 to 30 cycles from step 2 to step 4)
Step 5: 4°C hold

Notes:
Other cycle conditions may work well for individual users. These conditions work well for most of the samples processed in our lab using reduced volume and reduced terminator mix concentrations.