Source

ChlorellavirusIL-3A
Note: Purifiedfromarecombinantsource(PatentNo.US005472872A)

ReagentsSupplied

10XCviJI*ReactionBuffer
DMSO 

Description

• CviJI*isanuniquerestrictionenzymecapableofdigestingDNAattwoorthreebaserecognitionsequence(1,2)
• CviJI(Cat.No.2125-01)normallycleavesthesequence5"...PuGCPy...3"betweentheGandCtoleavebluntends
• Under"relaxed"conditions(inthepresenceofMg2+,ATPandenhancers),CviJI*cleavesthesequences5"...GC...3"except5"...PyGCPu...3"
• Capableofcleavingsingle-strandedDNAanddouble-strandedDNAintosmall20-200bpfragments
• GeneratesnumeroussequencespecificoligonucleotidesfromunknownDNAsamples
• ExclusivelyfromChimerx

Applications

• CviJIandCviJI*cleaveDNAextremelyfrequentlyandthuscanbeusedforavarietyofnovelmolecularBIOLOGyapplications(2,3,4)
• CviJI/CviJI*partialdigestscanalsobeusedinapplicationssuchasshot-guncloning,generatingquasi-randomlibraries(2)andepitopemappingorpanning
• CviJIdigestionofanonymousDNAproducesalargenumberofoligonucleotidesizedpolymersuponthermaldenaturation,whichcanbeexploitedinapplicationssuchas: 

1. Large-scalemappingorsequencingprojectsutilizinganonymousprimers
2. HighresolutionmapppingofshortDNAs
3. Nucleicacidlabeling(ThermalCycleLabeling,4,Fig.1)
4. Detection(5)
5. Amplification(5)
6. Cloning(2,3)
7. Captureofnucleicacids

IncubationTemperature

37°C

HeatInactivation

65°Cfor10minutes

AssayUnitSubstrate

pBR322

ReactionBuffer

10XCviJI*ReactionBuffer:
20mMglycyl-glycineKOH(pH8.5)
10mMmagnesiumacetate
50mMpotassiumacetate
0.1mMATP
0.1mMdithiothreitol
Note:100% DMSOsuppliedseparately

StorageBuffer

20mMTris-acetate(pH8.0at4°C)
0.5mMEDTA
0.1mMdithiothreitol
5mMmagnesiumchloride
50mMpotassiumacetate
50%(v/v)glycerol

AssayConditions

20mMglycylglycine-KOH(pH8.5)
10mMmagnesiumacetate
0.1mMdithiothreitol
50mMpotassiumacetate
0.1mMATP
30%DMSO
1µgpBR322
Incubationfor3hoursat37°Cinatotalreactionvolumeof25µl

StorageConditions

Storeat-80°C
Shippedondryice

CustomerNote(s)

(1)CviJI*restrictionendonucleaseisinhibitedbyglycerolconcentrationsinexcessof2.5%.ThereforetheextensionofthedigestiontimeisrecommendedratherthanusingadditionalunitsofCviJI*.Alternatively,DNAsamplecanbeethanolprecipitatedandre-digested.

(2)DuetoextremefrequencyofCviJI*/CviJIrecognitionsites,stericalinterferenceofcloselylocatedrecognitionsitesisobserved.Itresultsinslowerdigestionofsuchsites.Inconsequence,thegeneratedoligonucleotidefragmentsarerarelyshorterthan15bpthatmakesthemidealforanonymousprimerapplications.

(3)CviJI*reactionbuffercontainsDMSO,whichdoesnotinterferewithfurtherenzymaticmanipulations(ligations,labeling,etc).Ifthesampleisintendedforelectrophoresis,ethanolprecipitatationofthereactionmixtureaftercompleteddigestionisstronglyrecommendedinordertoavoiddiffusedbandsonagaroseorpolyacrylamidegels.

Downloads

CertificateofAnalysisPDF -CurrentLot
MSDSPDF -CurrentLot

References

(1)Xia,Y.,Burbank,D.,Uher,L.,Rabussay,D.andVanEtten,J.NucleicAcidsRes.15,6075-6090
(2)Fitzgerald,M.C.,Skowron,P.,VanEtten,J.L.,Smith,L.M.andMead,D.A.(1992)NucleicAcidsRes.20,3753-3762
(3)Skowron,P.M,Swaminathan,N.,McMaster,K.,George,D.,VanEtten,J.andMead,D.Gene157(1995)37-41
(4)Mead,D.,Swaminathan,N.,VanEttenJ.andSkowron,P.M.:RecombinantCviJIrestrictionendonuclease.(1995)UnitesStatesPatentnoUS005472872A
(5)Swaminathan,N.,McMaster,K.,Skowron,P.andMead,D.A.AnalyticalBiochemistry255(1998)133-141