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Team:Sumbawagen/Notebook/protocol10
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Revision as of 23:43, 20 January 2015
Team:Sumbawagen/Team
From 2014.igem.org
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
4. DNA sequencing data in .ab1 files were received by email, then analyzed using Bioedit program. For alignment, online CLUSTAL program was used.
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