TspDTISource

PurifiedfromEcolistrainthatcarriestheclonedtspDTRIgenefromThermusspeciesDT

UnitDefinition

Oneunitistheamountofenzymerequiredtocompletelydigest1 µgofpUC19DNAtoobtainastabledigestionpatternin1hourinatotalreactionvolumeof30 µl

StarActivity

N/A

ReactionTemperature

70°C

HeatInactivation

No

AssayUnitSubstrate

pUC19DNA

ReactionBuffer

10mMTris-HCl(pH8.5at25°C)
10mMMgCl₂
1mMdithiothreitol
+Enhancers(1)

Notes:

1. Avoidmultiplecyclesoffreezing/thawingofthestockreactionbuffer;Nomorethanthree[3]times.
2. Thawingshouldbepreformedattemperaturesnotexceeding10°C.
3. Recommendedprocedureistodividetheprovidedreactionbufferintosmallerportionsandpreservethemat-70°Cforlongtermstorage.
4. Recommendedshorttermstorageofreactionbufferisat    -20°C.

StorageBuffer

20mMTris-HCl(pH8.3at25°C)
25mM(NH₄)₂SO₄ 
25mMKCl
0.5mMEDTA
0.5mMdithiothreitol
0.02%TritonX-100™
0.02%Tween20™
0.02%Igepal
50%(v/v)glycerol

AssayConditions 

10mMTris-HCl(pH8.5at25°C)
10mMMgCl₂ 
1mMdithiothreitol 
1 µgpUC19DNA
+Enhancers(1)
Incubationisat70°Cfor1hourinareactionvolumeof30 µl 

Notes

1. Itisnotrecommendedtousemorethan2unitsper30 µlreaction.Itissuggestedtoperformdigestionforover1hour.
2. ToavoidDNAshiftduringelectrophoresiscausedbystrongprotein-DNAinteraction,itisrecommendedtoterminatereactionbyadditionofreactionstopsolution(containingdenaturingreagent,i.e.0.2%SDS)followedbyheatinactivationat89°Cfor20minutes.

StorageConditions

Storeat-20°C
Shippedondryice

References

(1)Skowron,P.M.,Majewski,J.,Zylicz-Stachula,A.,Rutokowska,S.M.,Jaworowska,I.,andHarasimowicz-Slowinska,R.I.(2003)NucleicAcidsResearch31,14e74

Downloads

CertificateofAnalysisPDF-CurrentLot
MSDSPDF-CurrentLot

Categories

RestrictionEndonucleases