GsuI (BpmI)
Detailní popis
Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
Highlights
- Superior quality – stringent quality control and industry leading manufacturing process
- Convenient color-coded Five Buffer System
- Includes universal Tango buffer for double-digestions
- BSA premixed in reaction buffers
- Wide selection of restriction endonuclease specificities
Note
- Incubation at 37°C results in 70% activity.
- GsuI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.01 mM S-adenosylmethionine gives a 2-fold increase in activity.
- GsuI may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.
- GsuI is blocked by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.
For methylation sensitivity refer to product specifications.
Conditions for 100% Activity |
|
Double Digestion | Perform double digestion using DoubleDigest. |
Hazardous | No |
Isoschizomers | Search for commercial isoschizomers using REsearch. |
Storage Buffer | GsuI is supplied in: 10 mM potassium phosphate (pH 7.5 at 25°C), 1 mM EDTA, 1 mM DTT, 0.2 mg/mL BSA, and 50% (v/v) glycerol. |
Storage Condition | -20 C |
Reaction conditions
Recommended buffer for 100% activity | Optimal temperature | Enzyme activity in Thermo Scientific buffers, % | Tango buffer for double digestion | |||||
---|---|---|---|---|---|---|---|---|
B (blue) 1X |
G (green) 1X |
O (orange) 1X |
R (red) 1X |
Tango (yellow) 1X / 2X |
||||
B (Blue) | 30°C | 100 | 50-100 | 0-20 | 20-50 | 100 | 50-100 | 1X or 2X |
Methylation Effects
- Dam: never overlaps – no effect.
- Dcm: may overlap – blocked.
- CpG: never overlaps – no effect.
- EcoKI: never overlaps – no effect.
- EcoBI: never overlaps – no effect.
Methylation type | Sequence | Cleavage effect |
---|---|---|
Dcm (CCWGG ) |
5'...CTCm5CA GG ...3'3'... GAG GTm5CC ...5' |
Blocked |
Cleavage efficiency close to the termini of PCR fragments
bp from the recognition site to fragment end | ||||
---|---|---|---|---|
1 | 2 | 3 | 4 | 5 |
50-100 |
Number of recognition sites in DNA molecules
Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
---|---|---|---|---|---|
25 | 3 | 2 | 4 | 1 | 1 |
pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
1 | 1 | 2 | 1 | 1 | 4 |
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