Applications

• Preferedsubstratesaresingle-strandedDNA,double-strandedDNAwith3’-hydroxylterminiandoligodeoxynucleotideprimers(1)
• Usedforspecificlabelingof3"-terminiwithribonucleotides(2)
• Labels3"-endsofDNAfragmentswithan[α-³²P]3"-deoxynucleoside(3)
• AddshomopolymertailsofdeoxyribonucleotidestovectorsorCDNAs(4,5)

ReagentsSupplied

5XTdtC-TailingReactionBuffer

UnitDefinition

Oneunitistheamountofenzymerequiredtotransfer1nmolofdAMPfromdATPtothe3"OHterminusoftheoligonucleotideinitiatord(A)15for1hourat37ºC

HeatInactivation

15minutesat 75ºC

Concentration

10-15units/µl

StorageBuffer

50mMpotassiumphosphate(pH7.4) 
1mMβ-mercaptoethanol
50%(w/v)glycerol

AssayConditions

40mMKCacodylate(pH7.2)
8mMMgCl₂
0.33mMZnSO₄
10µMoligonucleotided(A)₅₀
10µl bovineserumalbuminbuffer 
1mM(α-³²P)dATP
Reactionvolumeof60µl

Note:Allreactionsshouldberuninpolypropylenetubes

StorageConditions

Storeat-20°C
ProductshippedonDryIce

Downloads

CertificateofAnalysisPDF -CurrentLot
MSDSPDF -CurrentLot   

References

(1)Chang,L.M.andBollum,F.J.(1971)J.Biol.Chem.246,909-916
(2)Roychoudury,R.andKossel,H.(1971)Eur.J.Biochem.22.310-320
(3)Tu,C.P.D.andCohen,S.N.(1980)Gene10,177-183
(4)Roychoudhury,R.,Jay,E.andWu,R.(1976)NucleicAcidsRes.3,863-877
(5)Deng,G.andWu,R.(1983)MethodsEnzymol.100,96-116