• Highstranddisplacementactivity.
  • UsedinisothermalamplificationandNextGenerationSequencing.
FreeSampleAvailable

HighspecificactivityforDNAamplificationandsequencing.

Applications

  • Stranddisplacementamplification*
  • DNAsequencingthroughhighGCregions(1,2)
  • RapidsequencingfromnanogramamountsofDNAtemplate(3)

BstDNAPolymerase,ExonucleaseMinus,isarecombinantformofthe67kDaBacillusstearothermophilusDNAPolymeraseprotein(largefragment).Theenzymehas5’-3’polymeraseactivityandstranddisplacementactivity,butitlacks3’-5’exonucleaseactivity.Italsohasreversetranscriptionactivity.

Lucigen'sBstDNAPolymerase,ExonucleaseMinus,hashigherstranddisplacementactivitythanthatofothersuppliers(Figure1).Theenzymecanbeusedinnucleicacidamplificationmethods*suchasisothermalamplification,wholegenomeamplification(WGA),andmultipledisplacementamplification(MDA).ItalsocanbeusedinNextGenerationsequencing.

Thisenzymehasoptimalactivityat65°C.ItissuitableforsequencingDNAwithhighGCcontentandsecondarystructures.Itisavailableinconcentrationsof8,000U/mlor50,000U/ml.

BstDNAPolymerase,ExonucleaseMinus,issuppliedwith10XDNAPolymeraseBufferB,composedof200mMTris-HClpH8.8,100mM(NH4)2SO4,100mMKCl,20mMMgSO4,and1.0%TritonX-100.

Lucigen Bst versus competitors.

Figure1.LucigenBstDNAPolymerase,ExonucleaseMinus,possessesgreaterstrand-displacingpolymeraseactivity.M13singlestrandedDNAwasincubatedwithorwithout8unitsofBstDNAPolymerase(+/-Bst)inreactionbuffersuppliedbythemanufacturer,withorwithoutreplicationprimer(+/-primer)for30minutesat65°C.MW,1kbladder.

Tipsforuse:

  • Requires0.1%TritonX-100forlongtermstorage.
  • Reactiontemperaturesabove70°Carenotrecommended.
  • BstDNAPolymerasecannotbeusedforthermalcyclesequencing.

HeatInactivation:80ºCfor20min.

References:

  1. Griffin,H.andGriffin,A.(1994)PCRTechnology,228-229.
  2. McClary,J.etal.(1991)J.DNASequencingandMapping,1,173-180.
  3. Mead,D.A.etal.(1991)Biotechniques,11,76-87.


*Note:
Someusesforthisproductmayrequirelicenses.Lucigendoesnotencourageorsupporttheunauthorizedorunlicenseduseofpatentednucleicacidamplificationprocessesforisothermalamplification,wholegenomeamplification(WGA),multipledisplacementamplification(MDA),andNextGenerationsequencing.Itisthesoleresponsibilityofthebuyertoensurethatuseoftheproductdoesnotinfringethepatentrightsofthirdparties.Ifthepurchaserisnotwillingtoaccepttheseuselimitations,LucigenCorporationiswillingtoacceptreturnoftheproductforafullrefund.

ORDERINFORMATION

BstDNAPolymerase,ExonucleaseMinusisavailableintwoconcentrations,8,000U/mland50,000U/ml,inastoragebufferof10mM Tris-HCl,pH7.5,50 mM KCl,1 mM DTT,0.1 mM EDTA,0.1% TritonX-100,and50% Glycerol.Alsosuppliedis10XDNAPolymeraseBufferB,composedof200 mM Tris-HClpH8.8,100 mM (NH4)2SO4,100 mM KCl,20 mM MgSO4, and1.0 % TritonX-100.