View Single Post
Old 12-18-2018, 12:23 PM   #2
Junior Member
Location: Nevada

Join Date: Dec 2018
Posts: 2

So ONT contacted me with an answer to my question.
The Blunt/TA ligase master mix was validated in-house for the ligation of the barcodes. However, feel free to just use T4 from the Quick Ligation Module. If you go this route, please see the following suggestion:

So, the total reaction would include:

End prep DNA 22.5 Ál

Native Barcode Adapter 2.5 Ál

NEBNext Quick T4 DNA Ligase 10 Ál

NEBNext Quick Ligation Reaction Buffer (5X) 15 ul

Total 50 Ál

I plan on skipping the FFPE repair step because I plan preping fresh gDNA from bacteria cultures. Has anybody tried this before?
aharring is offline   Reply With Quote