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-   -   ABB buffer (http://seqanswers.com/forums/showthread.php?t=83196)

grace_pang 06-29-2018 11:48 AM

ABB buffer
 
Does anyone know what's in the ABB buffer?

I used Ligation Sequencing Kit 1D to prepare my sample. After ligation with the adapter using NEB blunt/TA kit, I used Ampure beads to clean the reaction. Normally when using Ampure beads, I wash it twice with 80% ethanol. But the ONT protocol says washing twice using ABB buffer. So I did it accordingly. But the library is not good, as I got ~ 300 recovering pores on the MinION flowcell. This indicates contamination in the library, right?

So is this ABB buffer important for the run? Has anyone washed the library with just 80% ethanol?

flokraft 07-12-2018 02:55 AM

It is a wash buffer without ethanol, because in the final wash step the sequencing adapters with the helicase are already attached to the DNA. With an ethanol washing step you would distroy and remove the helicase.

grace_pang 07-13-2018 08:00 AM

Thank you flokraft.


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