Notebook – Protocol – 10. DNA Sequencing

DNA sequencing was done by ordering to commercial company.

1. Premix sample was prepared with the following composition:

• Plasmid sample, 10 to 20 ul*
• 10 uM primer, 1 ul
• Sterilized distilled water, 9 to 19 ul
Total volume, 21 ul
Concentration < 900 ng

2. Samples were sent to the company

3. According to the company, DNA sequencing was done using the following conditions:

• Sequencing kit, BigDye® Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems)
• Purification kit, BigDye XTerminator® Purification Kit (Applied Biosystems)
• Sequencer, Applied Biosystems 3730xl DNA Analyzer

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