Sequencing DNA

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(Notes)
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==Notes==
==Notes==
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At least at the biopolymers facility and probably true for most sequencing results, when doing a run-off sequencing reaction, an extra (sometimes pretty high amplitude) 'A' peak is seen at the end before the template ends. The workers at the biopolymers facility seemed surprised when they were told about this, but they were able to find out that the sequenase enzyme used in the sequencing reaction is a genetically modified form of Taq. Therefore, it is most probable that the extra A is the template-independent A that Taq tends to add to the 3'-end of DNA.
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At the MIT biopolymers facility and probably with most sequencing centers, when doing a run-off sequencing reaction, an extra (sometimes pretty high amplitude) 'A' peak is seen at the end before the template ends. The workers at the biopolymers facility seemed surprised when they were told about this, but they were able to find out that the sequenase enzyme used in the sequencing reaction is a genetically modified form of Taq. Therefore, it is most probable that the extra A is the template-independent A that Taq tends to add to the 3'-end of DNA.

Revision as of 17:28, 3 December 2005

Specific Protocols

MIT:Sequencing BioBrick DNA -- protocol for DNA sequencing of BioBrick parts

Notes

At the MIT biopolymers facility and probably with most sequencing centers, when doing a run-off sequencing reaction, an extra (sometimes pretty high amplitude) 'A' peak is seen at the end before the template ends. The workers at the biopolymers facility seemed surprised when they were told about this, but they were able to find out that the sequenase enzyme used in the sequencing reaction is a genetically modified form of Taq. Therefore, it is most probable that the extra A is the template-independent A that Taq tends to add to the 3'-end of DNA.

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