Lambda Exonuclease (E. coli)

Source: CHIMERx

One unit produces 10 nmol of acid-soluble deoxribonucleotide from double-stranded DNA in 30 min at 37°C.

Details

For unambiguous PCR Sequencing: 1 Digest purified DNA with Recombinant Lambda Exonuclease 2 Heat to inactivate enzyme 3 Ready for DNA sequencing

Double-stranded specific DNase that requires the presence of a 5'-phosphate group for activity
Can be treated with lambda exonuclease to yield a single-stranded DNA when PCR products are prepared using one PCR primer phosphorylated at the 5'-end and the other primer unphosphorylated
When sequenced, the single-stranded DNA yields a clean readable sequence without the extraneous bands that are often present when PCR products are sequenced directly (Figure 2)
Effects are especially noticeable when sequencing PCR products with a high GC content

Assay Conditions:

67 mM glycine-KOH, pH 9.4, 2.5 mM MgCl2, 20 µg/ml sonicated [3H]-labeled Lambda DNA, 50 µg/ml BSA and lambda exonuclease in 50 µl for 30 min at 37°C.

Quality Control:

All preparations are assayed for contaminating nonspecific endodeoxyribonuclease and 3' exodeoxyribonuclease activities. Tested for linear DNA.

Concentration:

1-10 units/µl.

CHIMERx, 5520 W. Burleigh St., Milwaukee, WI 53210. Tel: 414-535-8585; Fax: 414-535-9508.

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