I would like to ask for some help with my project. My aim is to sequence several mitochondrial genomes. I decided to amplify every genome in 2 or 3 amplicons (which would be about 8-10kbp), and then sequence them on both strands using 454 technology.
I’m sorry for being very basic. But I’m a little bit confused. I decided to use NGS because overall it would be cheaper than Sanger sequencing (at least I hope so).
So this is how I understand the workflow: I amplify my genomes in 3 amplicons, then I nebulize my products in order to get fragments which would be appropriate for sequencing. Then I add adapters and MIDs (different MIDs for different genomes) and pool it together.
How do I know what surface of a plate I need in order to sequence my data with reasonable coverage?
In my lab, I’m able to amplify my DNA only, and I’m wondering if every service provider is able to do the rest.
Many thanks
I’m sorry for being very basic. But I’m a little bit confused. I decided to use NGS because overall it would be cheaper than Sanger sequencing (at least I hope so).
So this is how I understand the workflow: I amplify my genomes in 3 amplicons, then I nebulize my products in order to get fragments which would be appropriate for sequencing. Then I add adapters and MIDs (different MIDs for different genomes) and pool it together.
How do I know what surface of a plate I need in order to sequence my data with reasonable coverage?
In my lab, I’m able to amplify my DNA only, and I’m wondering if every service provider is able to do the rest.
Many thanks
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