Details
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:
- Avoidmultiplecyclesoffreezing/thawingofthestockreactionbuffer;Nomorethanthree[3]times.
- Thawingshouldbepreformedattemperaturesnotexceeding10°C.
- Recommendedprocedureistodividetheprovidedreactionbufferintosmallerportionsandpreservethemat-70°Cforlongtermstorage.
- 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
- Itisnotrecommendedtousemorethan2unitsper30 µlreaction.Itissuggestedtoperformdigestionforover1hour.
- 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
ebiomall.com
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希望同路人多多交流啊!
这些染料都非常成熟,光毒和淬灭都很低,当然要考虑到你采集图像时的显微镜参数。
比如calcein,常用的示踪,绿光(虽然这些颜色只是根据光谱加上去的伪彩),但要考虑你的实验过程中结合细胞结构,是否会伴随calcein的泄漏,就是荧光降低。
dri,还可以看膜啊
cfda也不错
bcef虽是ph指示,但你试验时不仅可观察细胞,还可看细胞ph变化,也行。
当然你或许还要结合其他方法,如细胞免疫化学等手段去双染或多染,都需要综合考虑染料之间特性。单独染一个染料,有点浪费,不如多染,数据和图像也好看些。现在流行细胞成像。

