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商品描述
Details
Source
PurifiedfromEcolistrainthatcarriestheclonedtspGWRIgenefromThermusspeciesGW
UnitDefinition
Oneunitistheamountofenzymerequiredtodigest1 µgofpBR322DNAtoobtainastabledigestionpatternin1hourinatotalreactionvolumeof50 µl
ReactionTemperature
70°C
HeatInactivation
No
AssayUnitSubstrate
pBR322DNA
ReactionBuffer
10mMTris-HCl(pH8.5at25°C)
1mMdithiothreitol
10mMMgCl2
+Enhancers(1)
Notes:
- Avoidmultiplecyclesoffreezing/thawingofthestockreactionbuffer;Nomorethanthree[3]times.
- Thawingshouldbepreformedattemperaturesnotexceeding10°C.
- Recommendedprocedureistodividetheprovidedreactionbufferintosmallerportionsandpreservethemat-70°Cforlongtermstorage.
- Recommendedshorttermstorageofreactionbufferisat-20°C.
StorageBuffer
10mMTris-HCl(pH7.5at25°C)
0.1mMEDTA
50mMNaCl
1mMdithiothrietol
100µg/mlbovineserumalbumin
50%(v/v)glycerol
AssayConditions
10mMTris-HCl(pH8.5at25°C)
10mMMgCl2
1mMdithiothreitol
1µgpBR322DNA
Incubationisat70°Cfor1hourinatotalreactionvolumeof50 µl
Notes
- Itisnotrecommendedtousemorethan2unitsper50 µlreaction.
- Digestionshouldbeperformedforovertwo[2]hours.
- AsTspGWIbindsDNAverytightly,excessamountofTspGWIaddedcanretardDNAmigrationonagel,eveninthepresenceofdenaturingagents.
- TspGWIisveryhighlystimulatedbythepresenceoftworestrictionsitesinoppositeorientation;Boththedistancebetweenrecognitionsequencesandtheirimmediateneighborhoodalsoaffectsthecleavageeffectiveness;Singlesitesubstratesarecleavedslowly.
StorageConditions
Storeat-20°C
Shippedondryice
References
(1)Zylicz-Stachula,A.,Harasimowicz-Slowinska,R.,Sobolewski,I.andSkowron,P.M.(2002)NucleicAcidsResearch30,7e33
Downloads
CertificateofAnalysisPDF-CurrentLot
Categories
RestrictionEndonucleases
