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View additional product information for Anza™ 30 Bsu15I - FAQs (IVGN0306)
11 product FAQs found
The product information sheet for an individual restriction enzyme will indicate any sensitivity it may have to methylation patterns. One Shot INV110 competent cells (Cat. No. C7171-03) are dam- and dcm-. Plasmid DNA isolated from these cells is not methylated by Dam methylase or Dcm methylase and can be digested with restriction enzymes that are sensitive to such methylation.
All Anza restriction enzymes contain BSA.
Enzymes may freeze during shipment on dry ice. This does not affect them as they have been tested and shown to be 100% active after three freeze-thaw cycles. For 24-48 hour delivery, enzymes may be shipped on ice pack as their performance is not affected by short exposure times at 4 degrees C.
For optimal results, we highly recommend exclusively using Anza restriction enzymes in double digestion as they all have 100% buffer compatibility. If a certain enzyme is not available in Anza format, but is available in FastDigest format or as a Thermo Scientific conventional restriction enzyme, it may be possible to perform a double digestion. For specific protocol recommendations, please contact our technical support with detailed information of the desired enzymes and DNA template.
When the next step following digestion is agarose gel electrophoresis, the Anza Red buffer can be used directly. The Anza Red buffer contains the necessary loading dye and density gradient for directly loading on to the agarose gel. The red dye does not interfere with digestion and helps reduce additional pipetting.
Use the clear Anza Buffer for applications that require analysis by fluorescence excitation, as the dyes in the Anza Red Buffer may interfere with some fluorescence measurements.
Anza restriction enzymes contain a number preceding the name. This number provides the option to organize the enzymes in freezers numerically or alphabetically. More common enzymes tend to have a lower number.
Star activity is the nonspecific cleavage of sequences that are similar to a recognition sequence, but not identical to the recognition sequence. This occurs when reaction conditions are suboptimal. Some restriction enzymes have more of a propensity for star activity than others. Factors that are included in suboptimal conditions that result in star activity include high glycerol concentration, high concentration of enzyme, prolonged incubation time, and use of the incorrect buffer.
Anza restriction enzymes in conjunction with the Anza buffer have been formulated to eliminate star activity when following the recommended protocol. Anza enzymes allow for the flexibility for complete digestion in as few as 15 minutes and longer reactions, up to 16 hours, can be safely digested without showing star activity.
- A unit is defined as the amount of enzyme required to completely digest 1 µg of substrate DNA in 50 µL of the reaction mixture in 1 hour at 37 degrees C.
- Anza restriction enzymes and buffer have been formulated such that 1 µL of Anza restriction enzyme cleaves 1 µg of substrate DNA in 15 minutes in Anza buffer.
Isoschizomers are restriction enzymes that have the same recognition sequence and the same specificity. The first restriction enzyme discovered is called the prototype and all subsequently identified enzymes that recognize that sequence are isoschizomers.
-Digestions in several PCR reaction buffers were tested and the Anza restriction enzymes fully digested DNA. However, not all commercially available buffers could be tested.
To obtain the best results for unpurified PCR products, we recommend the following reaction conditions:
- Up to 10 µL of PCR reaction mixture
- 1 µL of 10X Anza Buffer
- 1 µL of Anza restriction enzyme
- Up to 20 µL of H2O
- 37 degrees C, 15 min
If digested PCR product will be used for cloning, it is necessary to purify the PCR fragment prior to digestion to remove the active polymerase. Active thermophilic DNA polymerase will still be present in the PCR mixture if it is not removed by purification and may alter the ends of the cleaved DNA thereby reducing ligation efficiency.
- All Anza modifying enzymes are compatible with the Anza buffer.
- Anza Alkaline Phosphatase is supplied with the Anza buffer.
- The Anza T4 PNK Kit, Anza DNA End Repair Kit, and the Anza DNA Blunt End Kit include their own buffers which contain co-factors for full functionality of each. They all are compatible with the Anza Buffer.
- The Anza T4 DNA Ligase Master Mix comes with no additional buffer and is fully compatible with Anza buffer.