Anybody has tried to use NEB restriction enzymes in Thermo FastDigest Green Buffers? Does it still works?
All of our lab's REs are from NEB, I am thinking about buying some FastDigest green buffer for quick digestion verification work.
Alternatively, is there any loading dye we can add into NEB cutsmart buffer which does not affect the digestion reaction?
Thanks
Like what Jorg C. de Ruijter said, i have also experienced the same thing. Low efficiency but successful digestion
Generally you can use buffer / enzyme combinations from different companies. Just have a look on the recipe of the buffer and check it with the recommended one from the enzymes manufacturer. Often there is hardly any difference.
Hi..I also tried this and it works successful but efficiency might be low.
So here is a bit of a witches hack I have done in the past. I have used a combination of NEB and FastDigest Enzymes: it seems to work fine for me, in fact well I would say. Make sure to use FastDigest and HF enzymes of each, but use 2ul of HF enzyme rather then one, and increase slightly the concentration of the buffer, so like 1.1X.
So it would look something like this:
5ul of Buffer
1ul of Fast Digest Enzyme
2ul of NEB HF Enzyme
DNA and Water to 40ul.
Incubate for longer, like 1 hour instead of 10-15 as recommended at 37 degrees. Also, if you are using enzymes with close max efficiency temp (https://international.neb.com/tools-and-resources/usage-guidelines/activity-at-37c-for-restriction-enzymes-with-alternate-incubation-temperatures) then you can increase the temperature.
The thing is, the two companies have purposefully engineered their systems to be non-compatible, and Fischer has not even have posted the composition of its buffers so its really hard to understand why they are not compatible... its really a money grab. What I present to you is really black magic as I can not directly tell you why some of these adjustments work, but they did for me.
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