>
产品中心 >
Target_Expression >
Excellgen/Thermus aquaticus Muts DNA错配修复蛋白,6X His-Taq Muts/EG-41/500µ;G
ProductName | ThermusaquaticusMutSDNAmismatchrepairprotein,6XHis-Taq-Muts |
Size | 500µg |
Description | TheTaqMutSDNAmismatchproteinrecognizesheteroduplexDNAscontainingmispairedorunpairedbases.ThisMutsproteinbindsinvitrotoheteroduplexDNAscontainingmispairedorunpairedbasesoverawidetemperaturerangefrom4to70 °CandhasaThermostableATPaseactivity.ThisthermostableTaqMutSisactiveattemperaturebetween0to75°C.SinceTaqMutSefficientlybindsto1-4basesdeletion(orinsertion)andmismatchbasepairsofGT,CTandAG,itisusefulfordetectingthesemutations.MutationscanbedetectedinpolyacrylamidegelsoronasolidphasesuchasNiagaroseorbeadsormagneticNi-NTAparticles. |
Applications |
|
Source | E.coli |
FusionTag | 6XHistagatC-terminus |
PurificationMethod | FPLC |
Concentration | 1mg/ml |
Purity | ~95%asdeterminedbySDS-PAGE |
Accession# | AAC43637,TAU3311,U33117 |
GeneName | ThermusaquaticusMutSDNAmismatchrepairprotein |
MW | 92.8kDa |
ProteinSequence | MKIEHMEGMLKGEGPGPLPPLLQQYVELRDQYPDYLLLFQVGDFYECFGEDAERLARALG |
StorageBuffer | 20mMTris-HCl,pH8.0,250mMNaCl,0.1mMEDTA,1mMDTT,50%Glycerol |
ReactionBuffer | 100mMKCl,50mMTris-HCl,pH8.5,5~20mMMgCl2,0.1mMEDTA,1mMDTT,2%Glycerol,65°C |
Storage | -20to-80°C. |
Shipping | 4°Cordryice |
Protocol (example) | 1.AfterfirstroundPCR,purifyPCRfragmentsusingQiagenQIAquickPCRpurificationkitwithelutionindH2O. 2.DilutePCRproductto250ng/µlin10mMTris–HCl,pH7.8,50mMNaClandheatto95oCfor5minfollowedbycoolingat0.1oC/sto25oC. 3.Addbindingbuffer(20mMTris–HCl,pH7.8,10mMNaCl,5mMMgCl2,1mMDTTand5%glycerol)toannealedPCRproduct,adjustDNAconcentrationto11.5ng/µl,add6XHis-MutSdimersto950nM. 4.Incubatethemixtureatroomtemperaturefor10min,thenaddanequalvolumeofNi-NTAbeadspreequilibratedwith1Xbindingbuffer,andincubatefor30minatroomtemperature. 5.Gentlyspindownbeadsandsavesupernatantforsubsequentprocessing(secondroundPCR,cloningetc). |
References | 1.Protein-mediatederrorcorrectionfordenovoDNAsynthesis.NucleicAcidsRes.2004Nov23;32(20):e162. 2.CorrectingerrorsinsyntheticDNAthroughconsensusshuffling.NucleicAcidsRes.2005Mar30;33(6):e55. 3.MutSasatoolformutationdetection.ActaBiochimPol.2005;52(3):575-83.Epub2005Aug4. 4.OnetubemutationdetectionusingsensitivefluorescentdyeingofMutSprotectedDNA.NucleicAcidsRes.2000Apr15;28(8):E36. 5.RapidSNPdiagnosticsusingasymmetricisothermalamplificationandanewmismatch-suppressiontechnology.NatureMethods-4,257-262(2007)doi:10.1038/nmeth1007 |