Library preparation solutions. Meridian is a primary manufacturer of specialized high-quality molecular reagents and offers solutions to a wide range of industries to diagnose and treat diseases, discover new therapeutics or develop tests for environmental, food and cosmetic safety.
Solutions for Next Generation Sequencing (NGS) Democratizing NGS Sequencing Through Innovative Reagents
With the reduction in cost and increased portability of sequencing instruments, consumables, and data analysis tools, NGS is becoming more accessible, affordable, and user-friendly for a wider range of researchers, clinicians, and industries. Recent technological advances have opened the door to smaller and more compact sequencing instruments that have a reduced footprint, user-friendly interface, simplified workflow, and lower sample requirements. These advantages reduce the cost of performing NGS and increase its portability, enabling researchers to conduct sequencing experiments in various settings, including fieldwork and point-of-care diagnostics. However, a continuous challenge for NGS researchers has been the requirement of cold-chain shipping and storage for assay reagents. Ensuring the integrity of the cold chain throughout the supply chain logistics, especially in environments with unreliable electricity or limited refrigeration capacity, has resulted in high costs and logistical hurdles, hindering the global accessibility of NGS technology. Meridian’s glycerol-free, high-concentration NGS enzymes are designed for lyophilization which enables ambient-temperature stability, providing a sustainable alternative to cold-chain logistics. Eradicating the reliance on energy-intensive refrigeration and freezing for storage and shipping not only reduces carbon emissions but simplifies supply chain logistics, avoids packaging waste, and slashes transportation costs. The enzymes’ high-concentration format also meets the demands for miniaturization, often required in point-of-care devices. To further increase the accessibility of NGS reagents, Meridian has developed a Lyophilized NGS Enzymatic DNA Fragmentation Kit ( MDX224 ) and a Lyophilized NGS Library Preparation Kit ( MDX219 ). They are a ready-to-use, complete solution for sequencing-by-synthesis (SBS) protocols that can be shipped and stored at ambient temperature for a minimum of 12 months. The Lyophilized NGS Enzymatic DNA Fragmentation Kit consists of one tube containing lyophilized components for time-controlled enzymatic fragmentation, and the Lyophilized NGS Library Preparation Kit consists of three tubes containing lyophilized components for end-repair and A-tailing, ligation, and library amplification. Together they provide a straightforward solution for the NGS library preparation process that is fast, convenient and cost-effective. Meridian’s glycerol-free NGS library prep enzymes and lyophilized kit empower NGS assay developers to engineer the next generation of NGS diagnostic tests and workflows.
Meridian’s NGS Library Prep Solutions
Lyophilized NGS Library Preparation Kit | Includes end-repair/A-tailing, ligation, and library amplification
MDX219
Lyophilized NGS Enzymatic DNA Fragmentation Kit
MDX224
NGS Clean and Select Beads
NGS Library Quantification Kit
MDX041
MDX039
MDX200 Glycerol-Free T4 DNA Ligase (HC)
Glycerol-Free T4 DNA Polymerase (HC)
MDX207
Glycerol-Free DNA Pol I Klenow Fragment (HC)
Glycerol-Free Taq HS High Conc. DNA Polymerase
MDX208
MDX170
Glycerol-Free High-Fidelity Pfu (HC)
MDX206 Glycerol-Free T4 Polynucleotide Kinase (HC)
MDX203
Key Product Features
G lycerol-free enzymes are fully active after lyophilization and storage at ambient temperature for a minimum of 12 months
H igh-concentration format is ideal for assays requiring reduced volumes, such as miniaturized point-of-care devices
High-concentration format also allows for flexibility and maximization of the sample input quantity to increase assay sensitivity
Lyophilized Library Prep Kit contains all the components for end-repair and A-tailing, ligation, and library amplification in three separate tubes that are stable at ambient temperature for a minimum of 12 months
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Meridian’s Glycerol-Free NGS Library Preparation Solution Overview of library preparation by fragmentation and relevant Meridian products to perform each step Fragments with 5’ and 3’ overhangs are repaired to create blunt ends, these are then phosphorylated and A-tailed, leaving ends amenable to adapter ligation. Sequencing adapters are then ligated to create complete sequencing-ready library fragments.
5’ 3’
5’ 3’
5’ 3’
5’ 3’
5’ 3’
5’ 3’
FRAGMENTATION
STEP 1
5’
5’
3’
3’
5’ 3’
5’ 3’
Lyophilized NGS Enzymatic DNA Fragmentation Kit (MDX224) SEE PAGE 3
3’
5’
5’
3’
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5’
5’
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3’
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5’ overhang 3’ 3’
3’ overhang
5’ 3’ 5’ 3’ 5’ 3’
5’ 3’ 5’ 3’
3’
3’
5’
5’
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5’ overhang
5’ overhang 5’ overhang
3’ overhang 3’ overhang 3’ overhang
5’
5’
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5’ 3’
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3’ 3’
3’ 3’
5’ overhang
3’ overhang
5’ 3’
5’ 3’
5’ 3’
5’ 3’
5’ 3’
CREATE BLUNT ENDS
STEP 2
(3’ – 5’ exonuclease activity)
(Fill in 3’ – 5’ direction)
3’ 3’
5’ 5’
5’ 5’
3’ 3’
5’ 5’ overhang 5’ overhang 5’ overhang P 3’
3’ overhang 3’ overhang 3’ overhang 5’ 3’ 5’ 3’
Glycerol-Free T4 DNA Polymerase (HC) (MDX207) SEE PAGE 4 Glycerol-Free DNA Pol I Klenow Fragment (HC) (MDX208) SEE PAGE 4
5’
3’ 5’ 3’
(3’ – 5’ exonuclease activity) (3’ – 5’ exonuclease activity)
(Fill in 3’ – 5’ direction) (Fill in 3’ – 5’ direction)
5’ 3’
5’ 3’
5’
5’
3’
3’
5’ 3’ 5’ 3’
5’ 3’ 5’ 3’
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(3’ – 5’ exonuclease activity) (3’ — 5’ exonuclease activity)
(Fill in 3’ – 5’ direction) (Fill in 3’ — 5’ direction)
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P
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3’
3’
(3’ – 5’ exonuclease activity) (3’ – 5’ exonuclease activity) (3’ – 5’ exonuclease activity) 5’ 3’ 3’ 5’ P 5’ P
P 5’ (Fill in 3’ – 5’ direction) (Fill in 3’ – 5’ direction) (Fill in 3’ – 5’ direction) 5’ 3’
5’ 3’
5’ 3’
3’
3’
P
5’
PHOSPHORYLATION
STEP 3
A
Glycerol-Free T4 Polynucleotide Kinase (HC) (MDX206) SEE PAGE 5
3’
P P
5’ 5’
3’
3’
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P 3’
P 5’ 5’
3’ 3’
A
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A
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3’ 3’ P 5’ P 5’ P 5’ A- 5’ A 3’ 3’ A 3’ -T A -T
P 3’
-A 5’ 5’ 5’ 5’ 5’ A A
P P 3’ P
3’ 5’
P
A
-T
5’
3’ 3’ P
A-TAILING
STEP 4
A-
T- A 5’
P
Glycerol-Free Taq HS High Conc. DNA Polymerase (MDX170) SEE PAGE 6
Adaptor -A T- -A T-
3’ 3’
Adaptor
A A P 5’ A- 3’ A -T A-
P P
3’
Adaptor
Adaptor
Adaptor
Adaptor
-A
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-T -T A-
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Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adapter
Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adaptor Adapter
ADAPTER LIGATION
STEP 5
A- A-
T- T-
-A
Glycerol-Free T4 DNA Ligase (HC) (MDX200) SEE PAGE 6
-T
-A
T-
Glycerol-Free High-Fidelity Pfu (HC) (MDX203) SEE PAGE 7 STEP 6 LIBRARY AMPLIFICATION
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NGS SOLUTIONS CATALOG
FRAGMENTATION
STEP 1
Lyophilized NGS Enzymatic DNA Fragmentation Kit
Cat. # MDX224 RXNS POUCHES 8 Fragmentation Mix
Lyophilized NGS Enzymatic DNA Fragmentation Kit (MDX224) is stable at ambient temperatures and can be rehydrated at the point of use, reducing handling steps and supporting consistent performance. Fragmentation variability can impact library quality and sequencing performance. MDX224 offers time-controlled fragmentation, allowing users to produce consistent and adjustable fragment sizes without mechanical shearing instruments.
Product Highlights • Ambient-temperature storage and global shipping • Reliable performance without freeze-thaw cycles • Consistent, time-controlled fragmentation across protocols • Ready-to-rehydrate format enables easy integration into existing workflows
Robust Time-Dependent Enzymatic Fragmentation
TIME (min)
AVG. SIZE (bp)
PEAK (bp)
8 12 15 Time (min)
8 min 12 min 15 min
8
900
960
12
580
562
15
450
143
Libraries were constructed from 1 µ g of human DNA. A fragmentation reaction time titration was conducted with MDX224, at 37°C for 8-, 12- and 15-minutes incubation. Progressive DNA fragmentation was observed, using a TapeStation (Agilent) with increasing incubation time, as indicated by the shift from high molecular weight genomic DNA to lower molecular weight fragments on the electropherograms. The changes in fragment size distribution and overall DNA integrity demonstrate time- dependent degradation of genomic DNA by MDX224.
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STEP 2 CREATE BLUNT ENDS
Glycerol-Free T4 DNA Polymerase (HC)
Cat. # MDX207
Product Highlights • Blunting of DNA ends by “filling in” or “polishing” the ends of double-stranded DNA fragments, for cloning or generating NGS libraries • Sanger sequencing, to remove fluorescent dye-labeled nucleotides as they are incorporated into the growing DNA strand • Concentration 20 U/ µ L VOLUME REACTIONS 50 μ L 500 Rxns 500 μ L 5,000 Rxns
Glycerol-Free T4 DNA Polymerase (HC) (MDX207) is supplied at concentration 20 U/ μ L and is a template dependent DNA polymerase that catalyzes the addition of nucleotides to the 3’ end of a DNA strand, extending it in the 5’ to 3’ direction. It also possesses 3’ to 5’ exonuclease activity, allowing it to remove nucleotides from the 3’ end of DNA strands but lacks a 5’ to 3’ exonuclease activity. It can be used for generating blunt ends on any duplex DNA molecule and for labeling DNA by replacement synthesis. In NGS library preparation it can be used separately or as part of an end- repair mix, where it can blunt DNA ends of double-stranded DNA fragments for subsequent ligation of adapters.
No Loss of Activity After Lyophilization
(Left) Glycerol-Free T4 DNA polymerase (HC) (MDX207) was lyophilized in its reaction buffer, and the stability was tested in an accelerated stability study. The lyophilized MDX207 ( red ) was incubated at 37°C for 1 month and its DNA polymerization activity was tested against a non-lyophilized aliquot stored at -20°C ( blue ). Results suggest that the lyophilized MDX207 is active following accelerated stability tests with projected 12 months stability at ambient temperature.
Liquid | Lyophilized
Glycerol-Free DNA Pol I Klenow Fragment (HC)
Cat. # MDX208
VOLUME REACTIONS 200 μ L 10,000 Rxns 1 mL 50,000 Rxns
Glycerol-Free DNA Pol I Klenow Fragment (HC) (MDX208) is supplied at 50 U/ μ L and is a large N-terminal truncated protein fragment produced from E. coli DNA polymerase I, which retains polymerase and 3’-5’ exonuclease activity but has lost 5’-3’ exonuclease activity. It can be used to fill in 5’-protruding ends and is ideal for DNA blunting by 3’ overhang removal and fill-in of 5’ overhangs prior to adapter ligation in next-generation sequencing library preparation.
Product Highlights • DNA blunting for NGS adapter ligation • Labeling by fill-in of 5’-overhangs of dsDNA • Concentration 50 U/ µ L
Fast Kinetics with High Performance
No Loss of Activity After Lyophilization
Meridian | NEB | Thermo | Qiagen
Glycerol-Free DNA Pol I Klenow Fragment (HC) was used in a primer extension assay. Glycerol-Free DNA Pol I Klenow Fragment (HC) was used at 5 U final in reaction with 4 ng of template and Lyo-Ready Klenow Reaction Buffer and compared to other suppliers under the same conditions. The results demonstrate that Glycerol-Free DNA Pol I Klenow Fragment (HC) has a faster reaction rate and higher end fluorescence than most suppliers.
Liquid | Lyophilized
Accelerated stability studies of Glycerol-Free DNA Pol I Klenow Fragment (HC) at 37°C in liquid ( blue ) or lyophilized ( red ) format. The results demonstrate that Glycerol- Free DNA Pol I Klenow Fragment (HC) is active following accelerated stability tests with at least a projected 12 months stability at ambient temperature.
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NGS SOLUTIONS CATALOG
STEP 3 PHOSPHORYLATION
Glycerol-Free T4 Polynucleotide Kinase (HC)
Cat. # MDX206 VOLUME REACTIONS 25 μ L 500 Rxns 250 μ L 5,000 Rxns
Glycerol-Free T4 Polynucleotide Kinase (HC) (MDX206) is supplied at 20 U/ μ L and is used to phosphorylate the 5’ end of both DNA and RNA for subsequent ligation by catalyzing the transfer of the γ -phosphate from ATP to the 5’ hydroxyl of single-stranded and double-stranded DNA. Glycerol-Free T4 Polynucleotide Kinase (HC) is used in NGS library preparation, either separately or as part of an end-repair mix, where phosphates are added to the 5’-hydroxyl terminus of double-stranded DNA for subsequent ligation of adapters by T4 DNA Ligase.
Product Highlights • 5’ phosphorylation for
subsequent ligation for NGS • Removes 3’ phosphoryl groups from DNA and RNA • Concentration 20 U/ µ L
Fast Kinetics with High Performance
A
B
0.012
0.01
0.008
0.006
0.004
0.002
0
NEB
Meridian
NEB | MDX206
A) Increase in fluorescence over time and B) phosphorylation reaction rate of Glycerol-Free T4 Polynucleotide Kinase (HC) ( blue ) and NEB’s T4 Polynucleotide Kinase ( red ) at a concentration of 0.005 U/ μ L. The results demonstrate that the glycerol-free enzymatic activity of T4 Polynucleotide Kinase produced by Meridian is comparable to glycerol-containing enzymes from other suppliers.
No Loss of Activity After Lyophilization
Liquid | Lyophilized
Accelerated stability studies of Glycerol-Free T4 Polynucleotide Kinase at 37°C for two months in lyophilized ( red ) compared to the liquid ( blue ) format, stored at -20°C. The results demonstrate that Glycerol-Free T4 Polynucleotide Kinase is not affected by lyophilization and subsequent storage at ambient temperature, fully retaining its activity when rehydrated.
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STEP 4 A-TAILING
Glycerol-Free Taq HS High Conc. DNA Polymerase
Cat. # MDX170 VOLUME UNITS 20 μ L 1,000 500 μ L 25,000
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. Glycerol-Free Taq HS High Conc. DNA Polymerase can be used for 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.
Product Highlights • High concentration polymerase allowing for reduced volume in a miniaturized format • Can be used for addition of sequencing adapters during NGS library preparation • Concentration 50 U/ µ L
Liquid | Lyophilized
STEP 5 ADAPTER LIGATION
Glycerol-Free T4 DNA Ligase (HC) (MDX200) is a high concentration enzyme (50 U/ 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
1,000 Weiss Units 10,000 Weiss Units
20 μ L
200 μ L
• Repairs single-stranded nicks in duplex DNA, RNA and DNA/RNA hybrids • Joins double-stranded oligonucleotide linkers or adapters to DNA, for use in NGS library construction and cloning • Concentration 50 U/ µ L
Free of Detectable of Exo- and Endonuclease Activities
Hindlll-digested λ DNA
Supercoiled pUC19 DNA
100 U T4 DNA Ligase
No Ligase Control
100 U T4 DNA Ligase
100 U T4 DNA Ligase
No Ligase Control
100 U T4 DNA Ligase
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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NGS SOLUTIONS CATALOG
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 UNITS 100 μ L 500 10 mL
Product Highlights • High concentration Pfu
20,000
(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 lyophilization (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. 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
Lyo Day 7 at 37°C
Lyo Day 28 at 37°C
Wet Reference
Lyo Day 0
AVAILABLE AS AN ALL-INCLUSIVE KIT
Lyophilized NGS Library Preparation Kit
Cat. # MDX219-A RXNS
POUCHES
Our Lyophilized NGS Library Preparation Kit (MDX219) is designed to be straightforward 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 • Ready-to-use (includes enzymes for end-repair /A-tailing, ligation, and library amplification), just add fragmented DNA • Cost-effective, sustainable and accessible testing for NGS 8
1 / ER/dA, 2 / LIG
Cat. # MDX219-B RXNS POUCHES 8 1 / ER/dA, 2 / LIG, 3 / PCR
Comparison of Lyophilized and Liquid Library Preparation Kits
A
B
LIBRARY
AVG. MAPPED READS (%)
Lyophilized Reagents
99.79 ± 0.04
Liquid Reagents
99.79 ± 0.12
Comparison of libraries created using lyophilized and liquid formulations. A) shows GC content and B) shows mapping efficiency 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.
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ADDITIONAL NGS PRODUCTS
Meridian’s NGS Clean and Select Beads (MDX041) are paramagnetic SPRI beads designed for clean-up and size selection of DNA fragments or NGS libraries. Paramagnetic beads selectively bind DNA fragments based on the volume ratio of bead suspension and sample. They are delivering highly purified DNA fragments, efficiently removing contaminants, such as nucleotides, primers, adapters, enzymes, buffer additives and salts. NGS Clean and Select Beads
Cat. # MDX041
VOLUME 50 mL 500 mL
Product Highlights • Paramagnetic beads for clean-up and size selection • Tuneable size selection protocols for single- or double-sided selection • Compatible with manual and automated workflows
Left-Side and Right-Side Size Selection of DNA Fragments
For double-sided size selection, supernatant from a right-side selection step (which excludes the largest library fragments) is re-purified using a different ratio of beads in a left-side purification step to exclude the smallest library fragments.
Meridian’s NGS Library Quantification Kit (MDX039) is a qPCR-based assay for the exclusive quantification of adapter-ligated molecules, providing accurate quantification of Illumina ® NGS libraries. It contains pre-diluted standards to minimize pipetting errors, a P5 / P7 Illumina ® -specific primer mix and an optimized buffer for dilution of NGS library samples. NGS Library Quantification Kit
Cat. # MDX039
VOLUME REACTIONS 5 mL 500
Product Highlights • Suitable for quantification of all Illumina ® - compatible libraries • Accurate results in less than 90 minutes • Contains six pre-diluted DNA standards for rapid and reliable standard curve generation
qPCR-Based Quantification of NGS Libraries
6 pre-diluted DNA standards | Illumina ® ®
Standard curve generated from the amplifications of the 6 pre-diluted DNA standards ( blue , 10 pM to 100 aM) and 3 dilutions of an Illumina ® NGS library ( red , ten-fold series).
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NGS SOLUTIONS CATALOG
FEB | 2026
Ordering Information
Glycerol-Free NGS Solutions
Lyophilized NGS Library Preparation Kit
MDX219
Lyophilized NGS Enzymatic DNA Fragmentation Kit
MDX224
Glycerol-Free T4 DNA Polymerase (HC)
MDX207
Glycerol-Free DNA Pol I Klenow Fragment (HC)
MDX208
Glycerol-Free T4 Polynucleotide Kinase (HC)
MDX206
Glycerol-Free Taq HS High Conc. DNA Polymerase
MDX170
Glycerol-Free T4 DNA Ligase (HC)
MDX200
Glycerol-Free High-Fidelity Pfu (HC)
MDX203
Supporting NGS Products
NGS Clean and Select Beads
MDX041
NGS Library Quantification Kit
MDX039
High-Fidelity Pfu
MDX003
High-Specificity Pfu HS Mix
MDX006
Lyo-Ready T4 Ligase Buffer, 5x
MDX204
RNase Inhibitor (Glycerol-Free)
MDX200
55C MMLV-RT
MDX117
EMAIL info@meridianlifescience.com ORDERS orders@meridianlifescience.com meridianbioscience.com/lifescience
5171 Wilfong Road Memphis, Tennessee, USA 38134
PHONE +1 901-382-8716 FAX +1 901-333-8223
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