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www.
lucigen.
comMA266E-RibonucleaseR,E.
coliFeb20191RibonucleaseR,E.
coliCat.
No.
RNR072502www.
lucigen.
com1.
IntroductionRibonucleaseR(RNaseR)fromE.
coli,isamagnesium-dependent3′→5′exoribonucleasethatdigestsessentiallyalllinearRNAsbutdoesnotdigestlariatorcircularRNAstructures,ordouble-strandedRNAwith3′-overhangsshorterthan7nucleotides.
1,2MostcellularRNAswillbedigestedcompletelybyRNaseR,withtheexceptionoftRNAs,5SRNAandintronlariats.
The3′-tailsoflariatswillbetrimmedbyRNaseRtothebranchpointnucleotide,wherethereisa2′,5′-phosphodiesterlinkage.
Lariatsareproducedduringpre-mRNAsplicingofintronregionsandcanbeisolatedfromamixtureoftotalRNAbydigestionwithRNaseR.
TheMasterPureCompleteDNAandRNAPurificationKitandMasterPureYeastRNAPurificationKitsareidealforsuchtotalRNApreparations.
RNAisolatedinthiswaycanbeusedasatemplatetoproducelabeledcDNAwhichisthenusedasatargetformicroarrayscontainingpotentialintronsequencesorfortilingarrayscontainingoverlappingregionsofcompletechromosomesorgenomes.
ThecDNAproducedinthiswaywillnotbealinearrepresentationoftheintron,butthesequencescontainedinitwillbeintron-derived.
RNaseRisprovidedasa250Usize(20U/L;1μg/L)andissuppliedwitha10XRNaseRReactionBuffer.
2.
ProductDesignationsandKitComponentsProductKitSizeCatalogNumberReagentDescriptionPartNumbersVolumeRNaseR250UnitsRNR07250RibonucleaseR(20U/L)E0111-20D112.
5L10XRNaseRReactionBufferSS000769-D1250L3.
ProductSpecificationsStorage:Storeonlyat-20°Cinafreezerwithoutadefrostcycle.
StorageBuffer:RNaseRissuppliedina50%glycerolsolutioncontaining50mMTris-HCl(pH7.
5),100mMNaCl,0.
1mMEDTA,0.
1%TritonX-100and1mMdithiothreitol.
UnitDefinition:Oneunitconverts1μgofpoly-r(A)intoacid-solublenucleotidesin10minutesat37°Cin20mMTris-HCl(pH8.
0),100mMKCland0.
1mMMgCl2.
10XRNaseRReactionBuffer:is:0.
2MTris-HCl(pH8.
0),1MKCland1mMMgCl2.
Note:RNaseRrequireslow(0.
1-1.
0mM)magnesiumconcentrationsforactivity.
LowEDTAconcentrationsinsubstrateRNAsolutionscannegativelyaffectRNaseRactivity.
AdditionalMgCl2,upto1mMfinalconcentrationcanbeusedtocompensateforEDTAinthesubstrate.
Optimalactivityisat37°C.
QualityControl:RNaseRisfunction-testedinareactioncontainingamixtureoflinearandcircularizedRNAoligonucleotides.
OnlythelinearRNAisdigested.
ContaminatingActivityAssays:RNaseRisfreeofdetectableendoribonucleaseandDNaseactivities.
techsupport@lucigen.
com888-575-969534.
ApplicationsAlternativesplicingstudiesGeneexpressionstudiesIntroncDNAproductionIntronicscreeningofcDNAlibrariesIsolationofsplicingintermediatesandlariats5.
References1.
Suzuki,H.
etal.
,(2006)Nucl.
AcidsRes.
34(8)e63.
2.
Vincent,H.
A.
andDeutscher,M.
P.
,(2006)J.
Biol.
Chem.
281(40)29769.
EpicentreisatrademarkofIllumina,Inc.
and/oritsaffiliate(s)intheU.
S.
andothercountries,andisusedunderlicense.
MasterPureisatrademarkofLucigen.
TritonisaregisteredtrademarkofRohm&Haas,Philadelphia,Pennsylvania.
lucigen.
comMA266E-RibonucleaseR,E.
coliFeb20191RibonucleaseR,E.
coliCat.
No.
RNR072502www.
lucigen.
com1.
IntroductionRibonucleaseR(RNaseR)fromE.
coli,isamagnesium-dependent3′→5′exoribonucleasethatdigestsessentiallyalllinearRNAsbutdoesnotdigestlariatorcircularRNAstructures,ordouble-strandedRNAwith3′-overhangsshorterthan7nucleotides.
1,2MostcellularRNAswillbedigestedcompletelybyRNaseR,withtheexceptionoftRNAs,5SRNAandintronlariats.
The3′-tailsoflariatswillbetrimmedbyRNaseRtothebranchpointnucleotide,wherethereisa2′,5′-phosphodiesterlinkage.
Lariatsareproducedduringpre-mRNAsplicingofintronregionsandcanbeisolatedfromamixtureoftotalRNAbydigestionwithRNaseR.
TheMasterPureCompleteDNAandRNAPurificationKitandMasterPureYeastRNAPurificationKitsareidealforsuchtotalRNApreparations.
RNAisolatedinthiswaycanbeusedasatemplatetoproducelabeledcDNAwhichisthenusedasatargetformicroarrayscontainingpotentialintronsequencesorfortilingarrayscontainingoverlappingregionsofcompletechromosomesorgenomes.
ThecDNAproducedinthiswaywillnotbealinearrepresentationoftheintron,butthesequencescontainedinitwillbeintron-derived.
RNaseRisprovidedasa250Usize(20U/L;1μg/L)andissuppliedwitha10XRNaseRReactionBuffer.
2.
ProductDesignationsandKitComponentsProductKitSizeCatalogNumberReagentDescriptionPartNumbersVolumeRNaseR250UnitsRNR07250RibonucleaseR(20U/L)E0111-20D112.
5L10XRNaseRReactionBufferSS000769-D1250L3.
ProductSpecificationsStorage:Storeonlyat-20°Cinafreezerwithoutadefrostcycle.
StorageBuffer:RNaseRissuppliedina50%glycerolsolutioncontaining50mMTris-HCl(pH7.
5),100mMNaCl,0.
1mMEDTA,0.
1%TritonX-100and1mMdithiothreitol.
UnitDefinition:Oneunitconverts1μgofpoly-r(A)intoacid-solublenucleotidesin10minutesat37°Cin20mMTris-HCl(pH8.
0),100mMKCland0.
1mMMgCl2.
10XRNaseRReactionBuffer:is:0.
2MTris-HCl(pH8.
0),1MKCland1mMMgCl2.
Note:RNaseRrequireslow(0.
1-1.
0mM)magnesiumconcentrationsforactivity.
LowEDTAconcentrationsinsubstrateRNAsolutionscannegativelyaffectRNaseRactivity.
AdditionalMgCl2,upto1mMfinalconcentrationcanbeusedtocompensateforEDTAinthesubstrate.
Optimalactivityisat37°C.
QualityControl:RNaseRisfunction-testedinareactioncontainingamixtureoflinearandcircularizedRNAoligonucleotides.
OnlythelinearRNAisdigested.
ContaminatingActivityAssays:RNaseRisfreeofdetectableendoribonucleaseandDNaseactivities.
techsupport@lucigen.
com888-575-969534.
ApplicationsAlternativesplicingstudiesGeneexpressionstudiesIntroncDNAproductionIntronicscreeningofcDNAlibrariesIsolationofsplicingintermediatesandlariats5.
References1.
Suzuki,H.
etal.
,(2006)Nucl.
AcidsRes.
34(8)e63.
2.
Vincent,H.
A.
andDeutscher,M.
P.
,(2006)J.
Biol.
Chem.
281(40)29769.
EpicentreisatrademarkofIllumina,Inc.
and/oritsaffiliate(s)intheU.
S.
andothercountries,andisusedunderlicense.
MasterPureisatrademarkofLucigen.
TritonisaregisteredtrademarkofRohm&Haas,Philadelphia,Pennsylvania.
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