T4 DNA Ligase & Restriction Enzyme Expression - I am looking to save money! (Mar/18/2009 )
I am a graduate student and I am looking to save money for our lab we largely do lots of molecular biology so we go through common enzymes (Ligase + EcoRI/BamHI) at a high rate. I am looking for anyone who expresses their own enzymes rather than buys them from NEB or Promega. I am looking for a protocol so that I can do this myself - if anyone has had success expressing and utilizing their own T4 DNA ligase or any sort of restriction enzymes I would like to know how it was done and what equipment you used. Thanks in advance, I appreciate it.
joelgray on Mar 18 2009, 11:08 AM said:
For enzymes like EcoRI and BamHI, it isn't worth your time making your own supply. These two enzymes are one of the cheapest restriction enzymes that you can buy. Just buy them in bulk and you can cut your self deal with NEB or Promega.
You can probably save money in other aspects of the plasmid construction process. Ie recycle your 50ml fulcon tube, use glass tube to culture your bacteria, recycle the silicon column used to bind DNA (for minipreps or gel extraction)
Also remember that you can usually dilute the enzymes and incubate longer, especially when the RE comes to you at 10 - 20 units /ul.
Enzymatics sells these enzymes in bulk at low prices.
And don't forget to factor in the time that it will take you to do it yourself, although you are not getting paid to work, your time is valuable to the lab, and you may find that the time you waste is not worth the money you save. Not to mention the fact that all of the quality control is done for you when you purchase enzymes.