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Old 03-09-2016, 06:44 AM   #1
Debortoli
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Default Agilent Haloplex DNA Ligase test

We bought this kit for DNAseq from Agilent (Haloplex Target Enrichment System) to use in Illumina Miseq. Right now, we have DNA Ligase (that came with the kit), for 5 reaction (samples preparation). If we had another tube of DNA Ligase, we would be able to raise up to 36 reactions, since we still have the other reagents sufficient for 36 reactions...

I saw on Agilent's website that they sell two different types of DNA Ligase: The PFU DNA Ligase and the T4 DNA Ligase.

I would like to know if somebody already tried to use one of these DNA Ligase in the kit to verify if works?

Thanks in advance...
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Old 03-09-2016, 09:22 AM   #2
cmbetts
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You could make a pretty good guess based on the reaction temperature where the ligase is used. Pfu ligase is from a thermophile, so if the temperature is high, Pfu, low, T4. I'd probably guess offhand that it's T4. The next question is whether it's a special high concentration enzyme and/or special buffer with crowding reagents to increase reaction efficiency.
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Old 03-09-2016, 09:38 AM   #3
Debortoli
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Quote:
Originally Posted by cmbetts View Post
You could make a pretty good guess based on the reaction temperature where the ligase is used. Pfu ligase is from a thermophile, so if the temperature is high, Pfu, low, T4. I'd probably guess offhand that it's T4. The next question is whether it's a special high concentration enzyme and/or special buffer with crowding reagents to increase reaction efficiency.
Hmm...I see. Actually, that's something we've been thinking. If there is something inside the kit ligase besides the ligase, that doesn't have in the other two sold by them.
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