Source

ThermusThermophilus111

Applications

• SuitableforhightemperaturesynthesisofDNA(1)
• TthIIIReverseTranscriptasesynthesizesCDNAfromRNAtemplate(2)
• ResultsingreaterspecificityofprimerhybridizationandextensionofRNA
• Canreversetranscribeatelevatedtemperatures(2)
• MinimizesproblemswithstrongsecondarystructureofRNA
• UsedforefficientPCRofDNA,containingproblematicsecondarystructures
• ApplicabletoRT-PCR;thesameenzymeisusedforbothreversetranscriptionandfollowingamplificationofobtainedcDNAtemplate(2)
• ResistanttoamplificationinhibitorspresentintemplateDNAisolatedfromproblematicsamples(3,4)

ReagentsSupplied

10XTth111ReverseTranscriptaseReactionBuffer
MnCl₂ Solution 

UnitDefinition

Oneunitistheamountofenzymerequiredtoincorporate1.0nmoleTTPintoacidinsolublematerialin10minutesat50°C

AssayConditions

40mMTris-HCl(pH8.5)
1mMMnCl₂
10mMdithiothreitol
1mg/mlbovineserumalbumin
0.4mMpolyA(dT)
0.5mM(α-³²P)TTP@10μCi/ml 
Incubationisfor10minutesat50°C inatotalreactionvolumeof50µl

StorageBuffer

50mMTris-HCl(pH7.5)
0.1mMEDTA
5.0mMdithiothreitol
50%(v/v)glycerol
StABIlizers

StorageConditions

Storeat-20°C
ProductshippedonDryIce

Downloads

CertificateofAnalysisPDF -CurrentLot
MSDSPDF   

References

1.Wang,A.M.,Doyle,M.V.andMark,D.F.(1989)Proc.Natl.Acad.Sci.USA86,9717-9721
2.Myers,T.W.,Gelfanld,D.H.(1991)Biochem.30,7661-7666
3.Kather,H.L.andSchwartz,I.(1994)Biotechniques16,84-92
4.Poddar,S.K.,Sawyer,M.H.andConnor,J.D.(1998)J.Med.Microbiol.47,1131-1135