STEP 4 A-TAILING
Glycerol-Free Taq HS High Conc. DNA Polymerase
Cat #MDX170
VOLUME
UNITS
No loss of activity after lyophilization (Right) Glycerol-Free Taq HS High Conc. DNA Polymerase, was lyophilized with its reaction buffer and stored at 37°C for 1 month Lyophilized enzyme ( red ) was tested in PCR amplification and compared with the enzyme stored at -20°C, not lyophilized ( blue ). Results demonstrate that the Glycerol-free Taq HS is not affected by lyophilization and subsequent storage at ambient temperature, fully retaining its activity when rehydrated. generating 3’-dA-tailed DNA fragments as it has terminal transferase activity and naturally leaves a 3’ terminal adenine making it ideal for ‘sticky end’ ligation of sequencing adapters during NGS library preparation. Glycerol-Free Taq HS High Conc. DNA Polymerase (MDX170) is supplied as a blend of high‑performance glycerol‑free Taq DNA Polymerase with a Taq hot-start antibody. Glycerol-Free Taq HS High Conc. DNA Polymerase can be used for
Product Highlights • H igh concentration polymerase allowing for reduced volume in a miniaturized format. • C an be used for addition of sequencing adapters during NGS library preparation. • Concentration 50 U/ µ L 20 μ L
1,000 500 μ L 25,000
Liquid | Lyophilized
STEP 5 ADAPTOR LIGATION
Glycerol-Free T4 DNA Ligase (HC) (MDX200) is a high concentration enzyme (50U/ μ L) that catalyzes the formation of a phosphodiester bond between adjacent 5’ phosphate and 3’ hydroxyl groups in duplex DNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex DNA. Glycerol-Free T4 DNA Ligase (HC)
Cat #MDX200
VOLUME
UNITS
Product Highlights • Efficiently joins blunt and cohesive ends. • Repairs single-stranded nicks in duplex DNA, RNA and DNA/RNA hybrids.
1,000 Weiss Units 10,000 Weiss Units
20 μ L
200 μ L
Free of detectable of exo- and endonuclease activities
• Joins double-stranded oligonucleotide linkers or adaptors to DNA, for use in NGS library construction and cloning. • Concentration 50 U/ µ L
Hindlll-digested λ DNA
Supercoiled pUC19 DNA
100 U T4 DNA Ligase
100 U T4 DNA Ligase
No Ligase Control
100 U T4 DNA Ligase
100 U T4 DNA Ligase
No Ligase Control
(Left) Glycerol-Free T4 DNA Ligase (HC) is free of detectable exo– and endonuclease activities. Glycerol-Free T4 DNA Ligase (HC) was incubated of for 16 hours at 37°C with 1 µ g of HindIII-digested λ DNA (Lanes 2 and 3) and supercoiled pUC19 DNA (Lanes 5 and 6). The results show no detectable exo- and endonuclease activities that may affect the ligation efficiency of the enzyme.
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