R5547 FAQs for PacI RE-Mix®, Master Mixes, Intl, NEB
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PacI RE-Mix® FAQ

Q1: What are the advantages of using a RE-Mix Restriction Enzyme Master Mix?
Q2: For some restriction enzymes star activity can be a concern. Is this also applicable to the RE-Mix?
Q3: The bands on my gel are not as sharp as I would like. Is there a way to improve the sharpness?
Q4: Is it possible to use RE-Mix in double digests?
Q5: Can RE-Mix be used in overnight digestions?

Q1: What are the advantages of using a RE-Mix Restriction Enzyme Master Mix?

A1: RE-Mix master mixes provide a fast and simple way to perform restriction digests. The master mix contains all the elements necessary to perform restriction digests in 5-15 minutes. Just add DNA and water and incubate the reaction at the recommended temperature.


Q2: For some restriction enzymes star activity can be a concern. Is this also applicable to the RE-Mix?

A2: Star activity in general is not a concern with RE-Mix as the recommended incubation time is short, the amount of glycerol is not more than 5% and the restriction enzymes used in the RE-Mix are not known to be prone to star activity. However, as is true with any restriction enzyme, extended digests, high glycerol and high enzyme concentration can lead to star activity.


Q3: The bands on my gel are not as sharp as I would like. Is there a way to improve the sharpness?

A3: Fuzzy bands are commonly seen with restriction digests when you do not use a stop solution and may appear if your agarose gel or running buffer is old. Use of fresh buffer, a fresh gel or addition of stop dye with EDTA will improve resolution.


Q4: Is it possible to use RE-Mix in double digests?

A4: Yes, double digests with RE-Mix are possible. Please follow the
double-digest protocol.


Q5: Can RE-Mix be used in overnight digestions?

A5: Yes, Re-Mix works anywhere from 5-15 minutes to overnight. You choose the incubation time that is most convenient to you.