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Old 12-23-2016, 02:25 AM   #1
jrb71
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Location: UK

Join Date: Dec 2014
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Default Alternative to NEB Blunt/TA ligase for ONT Rapid Sequencing Kit

Has anyone tried using a different TA ligase for library preparation? We've got a ton of KAPA Hyper Prep reagents (KK8503) which as far as I can work out can like for like substitute for the NEB Blunt/TA ligase master mix (and the NEB End repair/dA-tailing master mix though this isn't needed in the rapid library preparation). It seems a waste of cash to buy in new enzymes which we already have by the bucket load!
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Old 12-23-2016, 02:30 AM   #2
gringer
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Yes, I ran a flow cell in someone else's lab using some random other ligase that they had lying round. They just mixed the ligase with a buffer in the required concentration, and it was loaded as normal. The run worked, although I didn't have any comparable TA/ligase runs to benchmark against.

With the tiny amount required, you probably don't need the buffer, and could possibly get away with just adding a little bit more of anything that is ligase-like.
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