| ProductName | ThermusaquaticusMutSDNAmismatchrepairprotein,N-6XHis-MBP-Taq-Muts |
| Size | 500µg |
| Description | TheTaqMutSDNAmismatchproteinrecognizesheteroduplexDNAscontainingmispairedorunpairedbases.ThisMutsproteinbindsinvitrotoheteroduplexDNAscontainingmispairedorunpairedbasesoverawidetemperaturerangefrom4to70 °CandhasaThermostableATPaseactivity.ThisthermostableTaqMutSisactiveattemperaturebetween0to75°C.SinceTaqMutSefficientlybindsto1-4basesdeletion(orinsertion)andmismatchbasepairsofGT,CTandAG,itisusefulfordetectingthesemutations.MutationscanbedetectedinpolyacrylamidegelsoronasolidphasesuchasNiagaroseoramylosebeads,ormagneticNi-NTAparticles. |
| Applications |
|
| Source | E.coli |
| FusionTag | 6XHistagandMBPtagatN-terminus |
| PurificationMethod | FPLC |
| Concentration | 1mg/ml |
| Purity | ~95%asdeterminedbySDS-PAGE
|
| Accession# | AAC43637,TAU3311,U33117 |
| GeneName | ThermusaquaticusMutSDNAmismatchrepairprotein |
| MW | 135.9kDa |
| ProteinSequence | 1MGSSHHHHHHGTKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFP 61QVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPI 121AVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYA 181FKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTING 241PWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDE 301GLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVI 361NAASGRQTVDEALKDAQTGTDYDIPTTENLYFQGHMEGMLKGEGPGPLPPLLQQYVELRD 421QYPDYLLLFQVGDFYECFGEDAERLARALGLVLTHKTSKDFTTPMAGIPLRAFEAYAERL 481LKMGFRLAVADQVEPAEEAEGLVRREVTQLLTPGTLLQESLLPREANYLAAIATGDGWGL 541AFLDVSTGEFKGTVLKSKSALYDELFRHRPAEVLLAPELLENGAFLDEFRKRFPVMLSEA 601PFEPEGEGPLALRRARGALLAYAQRTQGGALSLQPFRFYDPGAFMRLPEATLRALEVFEP 661LRGQDTLFSVLDETRTAPGRRLLQSWLRHPLLDRGPLEARLDRVEGFVREGALREGVRRL 721LYRLADLERLATRLELGRASPKDLGALRRSLQILPELRALLGEEVGLPDLSPLKEELEAA 781LVEDPPLKVSEGGLIREGYDPDLDALRAAHREGVAYFLELEERERERTGIPTLKVGYNAV 841FGYYLEVTRPYYERVPKEYRPVQTLKDRQRYTLPEMKEKEREVYRLEALIRRREEEVFLE 901VRERAKRQAEALREAARILAELDVYAALAEVAVRYGYVRPRFGDRLQIRAGRHPVVERRT 961EFVPNDLEMAHELVLITGPNMAGKSTFLRQTALIALLAQVGSFVPAEEAHLPLFDGIYTR 1021IGASDDLAGGKSTFMVEMEEVALILKEATENSLVLLDEVGRGTSSLDGVAIATAVAEALH 1081ERRAYTLFATHYFELTALGLPRLKNLHVAAREEAGGLVFYHQVLPGPASKSYGVEVAAMA 1141GLPKEVVARARALLQAMAARREGALDAVLERLLALDPDRLTPLEALRLLQELKALALGAP 1201LDTMKG |
| 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-MBP–MutSdimersto950nM. 4.Incubatethemixtureatroomtemperaturefor10min,thenaddanequalvolumeofamyloseresin(NEB)preequilibratedwith1Xbindingbuffer,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 |
