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蚂蚁淘在线 / 品牌中心 / Excellgen / Excellgen/Thermus aquaticus Muts DNA mismatch repair protein, C-6X His-MBP-Taq-Muts/EG-197/500 µg

Excellgen/Thermus aquaticus Muts DNA mismatch repair protein, C-6X His-MBP-Taq-Muts/EG-197/500 µg

价格
￥7900.00
货号：EG-197
浏览量：127
品牌：Excellgen
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商品描述
ProductNameThermusaquaticusMutSDNAmismatchrepairprotein,C-6XHis-MBP-Taq-Muts
Size500µg
Description

TheTaqMutSDNAmismatchproteinrecognizesheteroduplexDNAscontainingmispairedorunpairedbases.ThisMutsproteinbindsinvitrotoheteroduplexDNAscontainingmispairedorunpairedbasesoverawidetemperaturerangefrom4to70 °CandhasaThermostableATPaseactivity.ThisthermostableTaqMutSisactiveattemperaturebetween0to75°C.SinceTaqMutSefficientlybindsto1-4basesdeletion(orinsertion)andmismatchbasepairsofGT,CTandAG,itisusefulfordetectingthesemutations.MutationscanbedetectedinpolyacrylamidegelsoronasolidphasesuchasNiagaroseoramylosebeads,ormagneticNi-NTAparticles.

Applications
  • RemovemismatchDNA(errorcorrection)fromgenesynthesisreaction
  • Mutationdetectionandremoval
  • RapidisothermalSNPdetection
SourceE.coli
FusionTagMBPtagatN-terminusand6XHistagatC-terminus
PurificationMethodFPLC
Concentration1mg/ml
Purity

~95%asdeterminedbySDS-PAGE

Accession#AAC43637,TAU3311,U33117
GeneNameThermusaquaticusMutSDNAmismatchrepairprotein
MW135.9kDa
ProteinSequence1MGKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPD
61IIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYN
121KDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDI
181KDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTS
241KVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKP
301LGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVD
361EALKDAQTNSSSNNNNNNNNNNLGDDDDKLEVLFQGPHMEGMLKGEGPGPLPPLLQQYVE
421LRDQYPDYLLLFQVGDFYECFGEDAERLARALGLVLTHKTSKDFTTPMAGIPLRAFEAYA
481ERLLKMGFRLAVADQVEPAEEAEGLVRREVTQLLTPGTLLQESLLPREANYLAAIATGDG
541WGLAFLDVSTGEFKGTVLKSKSALYDELFRHRPAEVLLAPELLENGAFLDEFRKRFPVML
601SEAPFEPEGEGPLALRRARGALLAYAQRTQGGALSLQPFRFYDPGAFMRLPEATLRALEV
661FEPLRGQDTLFSVLDETRTAPGRRLLQSWLRHPLLDRGPLEARLDRVEGFVREGALREGV
721RRLLYRLADLERLATRLELGRASPKDLGALRRSLQILPELRALLGEEVGLPDLSPLKEEL
781EAALVEDPPLKVSEGGLIREGYDPDLDALRAAHREGVAYFLELEERERERTGIPTLKVGY
841NAVFGYYLEVTRPYYERVPKEYRPVQTLKDRQRYTLPEMKEKEREVYRLEALIRRREEEV
901FLEVRERAKRQAEALREAARILAELDVYAALAEVAVRYGYVRPRFGDRLQIRAGRHPVVE
961RRTEFVPNDLEMAHELVLITGPNMAGKSTFLRQTALIALLAQVGSFVPAEEAHLPLFDGI
1021YTRIGASDDLAGGKSTFMVEMEEVALILKEATENSLVLLDEVGRGTSSLDGVAIATAVAE
1081ALHERRAYTLFATHYFELTALGLPRLKNLHVAAREEAGGLVFYHQVLPGPASKSYGVEVA
1141AMAGLPKEVVARARALLQAMAARREGALDAVLERLLALDPDRLTPLEALRLLQELKALAL
1201GAPLDTMKGKLAAALEHHHHHH
StorageBuffer20mMTris-HCl,pH8.0,250mMNaCl,0.1mMEDTA,1mMDTT,50%Glycerol
ReactionBuffer100mMKCl,50mMTris-HCl,pH8.5,5~20mMMgCl2,0.1mMEDTA,1mMDTT,2%Glycerol,65°C
Storage-20to-80°C.
Shipping4°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

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