Enzymes for Molecular Biology: Enzymes for Molecular Biology

Applications

• Preferred substrates are single-stranded DNA, double-stranded DNA with 3’-hydroxyl termini and oligodeoxynucleotide primers (1)
• Used for specific labelling of 3"-termini with ribonucleotides (2)
• Labels 3"-ends of DNA fragments with an [α-³²P] 3"-deoxynucleoside (3)
• Adds homopolymer tails of deoxyribonucleotides to vectors or cDNAs (4,5)

Reagents Supplied

5X Tdt C-Tailing Reaction Buffer

Unit Definition

One unit is the amount of enzyme required to transfer 1 nmol of dAMP from dATP to the 3" OH terminus of the oligonucleotide initiator d(A)15 for 1 hour at 37ºC

Heat Inactivation

15 minutes at75ºC

Concentration

10-15 units/µl

Storage Buffer

50 mM potassium phosphate (pH 7.4)1 mM β-mercaptoethanol50% (w/v) glycerol

Assay Conditions

40 mM K Cacodylate (pH 7.2)8 mM MgCl₂0.33 mM ZnSO₄10 µM oligonucleotide d(A)₅₀10 µlbovine serum albumin buffer1 mM (α-³²P) dATP Reaction volume of 60 µl

Note: All reactions should be run in polypropylene tubes

Storage Conditions

Store at -20°C

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200