STEP 6 LIBRARY AMPLIFICATION
Glycerol-Free High-Fidelity Pfu (HC) (MDX203) is a high-fidelity, thermostable DNA-dependent DNA polymerase from Pyrococcus furiosus . The 3’ to 5’ proofreading exonuclease activity of Pfu polymerase makes it suitable for applications that require high-fidelity DNA synthesis or blunt-ended PCR fragments. Glycerol-Free High-Fidelity Pfu (HC) is supplied in a glycerol-free storage buffer and is accompanied by a 5x Reaction Buffer that contains magnesium, dNTPs and excipients required for lyophilization. Glycerol-Free High-Fidelity Pfu (HC)
Cat #MDX203
VOLUME 100 μ L
UNITS
Product Highlights • High concentration Pfu
500 10 mL 20,000
polymerase allowing for bulk production of PCR master mixes. • Ideal for NGS library amplification. • Glycerol-free, ideal for preparation of dried-down PCR tests • Concentration 20 U/ µ L
Maintains multiplexing performance after lyophilization
(Left) Glycerol-Free High-Fidelity Pfu (HC) (MDX203), stored at 37°C for 1 and 4 weeks. Lyophilized samples were tested against the same enzyme and reaction buffer without lyophilisation (Wet Reference) in multiplexed PCR amplification of 7 targets of various GC content: 793 bp (32% GC), 649 bp (54% GC), 548 bp (60% GC) , 418 bp (48% GC), 332 bp (48% GC), 196 bp (51% GC), 135 bp (55% GC). Results show that the lyophilized MDX203 is stable with no loss of activity for at least 1 month at 37°C with projected ambient temperature stability of 1 year.
AVAILABLE AS AN ALL-INCLUSIVE KIT
Lyophilized NGS Library Preparation Kit
Cat #MDX219-A
REACTIONS
POUCHES
Our Lyophilized NGS Library Preparation Kit (MDX219) is designed to be straight forward and demonstrate the use of lyophilization in NGS library preparation. It is a ready-to-use complete solution for sequencing-by-synthesis (SBS) protocols, consisting of an end repair/dA-tailing mix module to convert fragmented DNA into 5’-phosphorylated and 3’-dA-tailed DNA fragments, a ligase module, for the addition of sequencing adapters and an optional high-fidelity PCR module for library amplification.
Product Highlights • Lyophilized reactions in sealed pouches for ambient temperature stability, transport and storage
8
1/ ER/dA, 2/ LIG
Cat #MDX219-B
REACTIONS
POUCHES 8 1/ ER/dA, 2/ LIG, 3/ PCR
• Ready-to-use (includes enzymes for end-repair/A-tailing, ligation, and library amplification), just add fragmented DNA • C ost-effective, sustainable and accessible testing for NGS
Comparison lyophilized and liquid library preparation Kits
A
B
Library
Average mapped reads (%)
Comparison of libraries created using lyophilized and liquid formulations. A) shows mapping efficiency and B) the GC content of the libraries generated with lyophilized and liquid reagents. In both cases lyophilized and liquid formulations showed similar results, with good mapping efficiency and comparable GC content.
Lyophilized reagents 99.79 ± 0.04
Liquid reagents
99.79 ± 0.12
6
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