Description | ATPindependentT4RNALigase2specificallyligatesthepreadenylated5´endofDNAorRNAtothe3´OHendofRNA.TheenzymedoesnotrequireATPforligationbutrequirespre-adenylatedlinkers. |
Applications | - Ligateapre-adenylatedDNAorRNAsequencetagtoanyRNA3´-end
- JoinasinglestrandedadenylatedprimertosmallRNAsforCDNAlibrarycreation
- JoinasinglestrandedadenylatedprimertoRNAforstrand-specificcDNAlibraryconstruction.
|
Source | PurifiedfromarecombinantE.colistraincarryingamutantofT4RNALigase2. |
Components | - ATPindependentT4RNALigase2,200,000units/mlinstoragebuffer:10mlTris-HCl,100mMNaCl,0.1mMEDTA,0.1mMDTT,50%Glycerol,pH7.5@25°C
- 10XT4RNALigase2truncatedBuffer:500mMTris-HCl,100mMMgCl2,10mMDTT,pH7.5@25°C
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UnitDefinition | 200unitsisdefinedastheamountofenzymerequiredtogive80%ligationofa31-merRNAtothepre-adenylatedendofa17-merDNAinatotalreactionvolumeof20μlin1hourat25°C. 5´-FAM-rArGrUrCrGrUrArGrCrCrUrUrUrArUrCrCrGrArGrArUrUrCrArGrCrArArUrA-3´ 5´-rAppCTGTAGGCACCATCAAT–NH2-3´ |
QualityControl | Theabsenceofendo-,exodeoxyribonucleases,phosphataseandribonucleasesconfirmedbyappropriatequalitytests. |
Storage | -20°C. |
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