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Old 12-18-2018, 12:23 PM   #2
aharring
Junior Member
 
Location: Nevada

Join Date: Dec 2018
Posts: 2
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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?
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