p50-T4 DNA Ligase Plasmid – Brought to you by Chemically Incompetent!

Published: February 5th, 2020   Last Modified: April 14th, 2020

One of the perks of running this site is meeting all sorts of interesting characters around the globe. Now and again you meet someone who really aligns with your values and one such fellow is Mark, who runs Chemically Incompetent. I highly encourage everyone to check out his website, he does an excellent job of blending science, philosophy and humor in his writing. If you like saucy lab hacks and ways to improve your day to day efficiency, then it’s right up your alley.

Mark has been working on a plasmid which encodes an improved version of T4 DNA Ligase fused to the p50 domain. This fusion is an upgrade over the venerable T4 Ligase and should save you heaps if you do massive amounts of cloning or you make your own Gibson mix.

He’s done a lot of work on the writeup and purification protocol, it’s clean and thorough. I’m happy to announce that he has allowed me to carry the plasmid as part of my plasmid pack. I will add my take on the purification protocol as soon as I give it a whirl! Thanks Mark! 🙂

5 thoughts on “p50-T4 DNA Ligase Plasmid – Brought to you by Chemically Incompetent!”

  1. Hello Mark, I just found your website. Really great !!!

    Do you know anyone who has tested the plasmid pCA24N-P50-ligase before? My graduate students and the rest of my team use a lot of ligase and it’s starting to get expensive !! I’m looking for an elegant way to produce it and saw that this plasmid is available. The purification is done with a nickel resin NTA.

    (sorry for my bad english (i’m french speaking)


  2. Dear sir,
    I need to ask the purified ligase is not working even after following the protocol. Please mention the probable reasons and their solutions.

    1. It looks like there may be a mutation in the sequence, it’s only visible from the 3′ end, so you’d have to sequence it with T7 term according to Ye. I had only sequenced it with T7 promoter so I never saw it. Please accept my apologies. I have the sequence for the mutagenesis primers to fix the plasmid if you’d like.

      1. Hello,
        I doubt how could the plasmid sequence get mutated? Is there some other reason during purification?
        If possible at your end, please do share your protocol you followed to purify the p50 ligase.

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