solutions for oligonucleotides synthesis liquid reagents from safc supply solutions®
DESCRIPTION
Purity of Liquid Reagents to support your overall synthesis performanceIn solid-phase synthesis of oligonucleotides, liquid reagents are used in each step of the synthesis cycle. Overall synthesis performance, and therefore total product yield and purity of the crude oligonucleotide, is highly dependent on the chemical purity of the monomers and the supporting liquid reagents.TRANSCRIPT
ProClin® Preservatives Mechanisms and Stability
for Diagnostics Reagents
www.safcsupplysolutions.com
Solutions for Oligonucleotides SynthesisLiquid Reagents
from SAFC Supply Solutions®
Solutions for Oligonucleotides SynthesisLiquid Reagents
2
SAFC Supply Solutions® has more than 25 years of manufacturing experience in DNA and RNA synthesis reagents. Our liquid reagents meet the most stringent quality specifications, making them ideal for use in automated nucleic acid synthesis. Lot-to-lot consistency is guaranteed for every product manufactured at SAFC Supply Solutions ISO 9001 certified site and each lot is quality-controlled to ensure optimum synthesis performance.
Chemical composition and purpose
Deblock solution containing trichloroacetic acid (TCA) or dichloroacetic acid (DCA) in dichloromethane or toluene removes the dimethoxytrityl (DMT) protecting group from the 5’hydroxyl moiety of nucleotides
already incorporated into the growing nucleic acid, prior to the addition of the next phosphoramidite. Removal of the DMT allows the unprotected 5’hydroxyl moiety to react with a new phosphoramidite in a subsequent extension reaction.
Wash solution containing anhydrous acetonitrile (water content ≤30 ppm), is used as the overall wash solvent following each step in the synthesis cycle.
Activator solution containing powerful activators like 4,5-dicyanoimidazole (DCI) or 5-(3,5-bis(trifluoromethyl) phenyl)-1H-tetrazole (Activator 42) in acetonitrile are mixed with solutions of phosphoramidites during the extension step. The activator reacts with the amidite group to form a highly reactive intermediate. The intermediate forms an internucleotide bond with the detritylated 5’hydroxyl group of the growing oligonucleotide chain.
Cap A solution containing acetic anhydride and tetrahydrofurane (THF) is used after the phosphoramidite reaction/coupling step. This solution aborts chains bearing unreacted 5’hydroxyl groups due to failure of reaction with activated phosphoramidite (typically, 1 to 2% of growing oligonucleotide chains in each synthesis cycle). These unreacted 5’hydroxyl groups are capped with an acetyl group, thus rendered unreactive for subsequent synthesis steps. For synthesis with Proligo® Reagents’ TAC-protected phosphoramidites, Fast Deprotection CAP A solution must be employed instead of Cap A solution.
Purity of Liquid Reagents to support your overall synthesis performanceIn solid-phase synthesis of oligonucleotides, liquid reagents are used in each
step of the synthesis cycle. Overall synthesis performance, and therefore total
product yield and purity of the crude oligonucleotide, is highly dependent on
the chemical purity of the monomers and the supporting liquid reagents.
Features Benefits
Products designed for oligonucleotide synthesis
Constant high synthesis performance
Liquid reagents offered in addition to amidites and supports
Key oligonucleotide synthesis reagents available in one shop
Constant high control
Matching with high quality amidites and support products
Broad range of packaging
Offering high flexibility
Customized formulations and specifications
Special requirements will be fulfilled
Technical expertise Supporting customized solutions
New product development: Activator® 42
Synthesis optimization and cost reduction
Table 1. Liquid Reagent features and benefits
Solutions for Oligonucleotides SynthesisLiquid Reagents
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Fast Deprotection Cap A solution must be used for synthesis with Proligo Reagents’ TAC-protected phosphoramidites. Fast Deprotection Cap A solution, with tert-butylphenoxyacetyl acetic anhydride (TAC2O) in tetrahydrofuran is used in place of Cap A solution to ensure that the displacement of tert-butylphenoxyacetyl (TAC) on guanine bases of the TAC-protected phosphoramidites does not occur.
Cap B solution containing tetrahydrofurane (THF) pyridine and N-methylimidazole (NMI),
is mixed in situ with Cap A solution during the capping (acetylation) reaction. Pyridine is applied as a mild base while NMI provides a powerful acylation catalyst.
Oxidizer solution, containing tetrahydrofurane (THF), pyridine, iodine and water is applied to oxidize the chemically unstable trivalent phosphorous triester linkage formed in the coupling reaction to a stable pentavalent phosphate triester. Iodine functions as a mild oxidant while water serves as an oxygen donor.
The Oligonucleotide Synthesis Cycle
Solutions for Oligonucleotides SynthesisLiquid Reagents
4
A broad range of packaging
Flexible packaging options are available to meet most synthesizer requirements, including ABI® 394, ABI® 3900, ExpediteTM, ÄKTATM and Mermade. As a single source supplier, SAFC Supply Solutions’ Hamburg (Germany) and Sheboygan, Wisconsin manufacturing sites can liquid fill and blend material in and have complete packaging
operations that can accommodate small-scale bottles to 1,400 L returnable containers.
Our worldwide distribution network offers advantages of dedicated airfreight space, expertise in chemical transport regulations and local delivery knowledge.
BottlesDescription Height Diameter Capacity Total Capacity Material Closure
100 mL Septum 95 mm 51 mm 100 mL 120 mL amber glass septum
4 oz 22-400 Bottle 112 mm 48,5 mm 120 mL 125 mL amber glass screw cap 22-400
8 oz 24-400 Bottle 137 mm 60 mm 200 mL 240 mL amber glass screw cap 24-400
8 oz 28-400 Bottle 137 mm 60 mm 200 mL 240 mL amber glass screw cap 28-400
16 oz 28-400 Bottle 170 mm 72 mm 450 mL 480 mL amber glass screw cap 28-400
32 oz 28-400 Bottle 210 mm 94 mm 900 mL 960 mL amber glass screw cap 28-400
2 L 38-430 Bottle 310 mm 135 mm 2 L 2.5 L amber glass screw cap 38-430
2.5 L GL 45 Bottle 296 mm 135 mm 2.5 L 2750 mL amber glass screw cap GL 45
4 L 38-430 Bottle 353 mm 161 mm 4 L 4300 mL amber glass screw cap 38-430
1 L GL 45 Bottle 212 mm 101 mm 1000 mL 1158 mL amber glass screw cap GL 45
100 mL Septum 4 oz 22-400 Bottle 8 oz 24-400 Bottle 8 oz 28-400 Bottle 16 oz 28-400 Bottle
32 oz 28-400 Bottle 2 L 38-430 Bottle 2.5 L GL 45 Bottle 4 L 38-430 Bottle 1 L GL 45 Bottle
Table 2. Worldwide Packaging Options
Solutions for Oligonucleotides SynthesisLiquid Reagents
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Drums
Description Capacity Total Capacity Weight Height Diameter Wall
Thickness Material Closure Operating Pressure
Approval Number
18 L Returnable Drum
Stainless steel drum 18 L with Dip Tube
18 L 20 L 6,6 kg 442 mm 278 mm 1.5 mm V2A 1.4301 SS304
Screw cap DIN 11851 PTFE R52x1/6”
0.5 bar 1A1/X2,0/900
25 L Drum, Disposal
Composite drum sheet steel/PE
25 L 30 L 3.4 kg 560 mm 280 mm 0.5 mm PE-HD A BCS 56x4 (1x) 1A1/X1,6/250
25 L Returnable Drum
Stainless Steel Drum with Dip Tube
25 L 30 L 10 kg 400 mm 380 mm 1.5 1.4301 3A Coding MAV Micro Matic Valve
0.5 bar 1A1/X2,3/900
45 L Returnable Drum
Stainless steel drum 45 L with Dip Tube
45 L 50 L 11 kg 600 mm 363 mm 1.5 mm V2A 1.4301 SS304
Screw cap DIN 11851 PTFE R52x1/6”
0.5 bar 1A1/X2,3/900
45 L Returnable Drum
Stainless Steel Drum with Dip Tube
45 L 50 L 12 kg 600 mm 263 mm 1.5 mm 1.4301 4F Coding MAV Micro Matic Valve
0.5 bar 1A1/X2,3/900
180 L Drum, Disposal
Composite drum sheet steel/PE
180 L 200 L 21 kg 884 mm 585 mm 1 mm PE-HD BCS 56x4 (2x) 1A1/X1,3/250
200 L Returnable Drum, Level Sensor
Stainless steel drum 200 L with level sensor and Dip Tube
200 L 225 L 43 kg 970 mm 600 mm 2 mm 1.4301 On Demand 0.5 bar 1A1W/X2,0/900
200 L Returnable Drum, Standard
Stainless steel drum 200 L with Dip Tube
200 L 225 L 43 kg 970 mm 600 mm 2 mm 1.4301 Screw cap DIN 11851 PTFE R52x1/6”
0.5 bar 1A1W/X2,0/900
Returnable 1400 L Container
Stainless Steel IBC 1400 L with Dip Tube
1400 L 1500 L 247 kg 1960 mm 1200 mm 1.4301 On demand 0.5 bar 31A/Y
18 L Returnable Drum 25 L Drum, Disposal 25 L Returnable Drum 45 L Returnable Drum 45 L Returnable Drum
180 L Drum, Disposal 200 L Returnable Drum, Level Sensor
200 L Returnable Drum, Standard
Returnable 1400 L Container
Table 3. Liquid Reagent Packaging
Solutions for Oligonucleotides SynthesisLiquid Reagents
6
For special requirements on formulations,
specifications and packaging, our manufact-
uring sites offer custom blending services:
• Batch sizes from 100 L up to 4000 L
• A wide variety of high purity solvents, including flammable solvents handling
• Raw materials supplier qualification process in alignment with customer expectations
• Specifications and Quality Control according to customer requirements
Six additional production plants support the Sheboygan and Hamburg facilities with custom blending and risk mitigation for all projects.
All inquiries are evaluated by a Project Management Group that selects the most appropriate manufacturing site and follows up on customer projects.
Custom Blending Solutions to speed up your processes
Description Capacity Total Capacity Weight Height Diameter Wall
Thickness Material Closure Operating Pressure
Approval Number
20 L Pure-Pac II 20 L 22 L 8 kg 20 inches 11 inches 1.5 mm SS304 MAV Micro Matic Valve - Simriz 485
0.34 bar 1A1/X2.3/900
50 L Pure-Pac II 50 L 55 L 15 kg 23 inches 16 inches 1.5 mm SS304 MAV Micro Matic Valve - Simriz 485
0.34 bar 1A1/X2.0/600
200 L Pure-Pac II 200 L 220 L 47 kg 38 inches 24 inches 2 mm SS304 MAV Micro Matic Valve - Simriz 485
0.34 bar 1A1/X2.0/600
200 L Pure-Pac II, level sensor
200 L 220 L 47 kg 38 inches 24 inches 2 mm SS304 MAV Micro Matic Valve - Simriz 485
0.34 bar 1A1/X2.0/600
Additional Drum Options (US market only)
For your custom packaging requirements, including technical and product specifications, please contact your sales representative to inquire. Some packaging options may not be available in your region.
Table 4. Liquid Reagent Packaging
Solutions for Oligonucleotides SynthesisLiquid Reagents
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Reagent Description Packaging Available
Formulation Specifications Product Number Unit Bottle Type
Amidite Diluent & Acetonitrile
ACN Water Content ≤ 30ppm or ≤10ppm also available *
L010010-06 6 x 100 mL 100 mL septum
nD20 = 1,342-1,346 L010200-04 4 x 2 L 2 L 38/430
App. of Sol. = clear L010250-04 4 x 2.5 L 2.5 L GL45
L010400-04 4 x 4 L 4 L 38/430
L011800-01 1 x 18 L 18 L Returnable Drum
L014500-01 1 x 45 L 45 L Returnable Drum
L014500-C01-01 1 x 45 L 45 L Returnable Drum
L0120000-01 1 x 200 L 200 L Returnable Drum 2” Level Sensor
L0120000-C01-01 1 x 200 L 200L Returnable Dru m 2” Standard
L0114000-01* 1 x 1400 L Returnable 1400L Container
L810045-06 6 x 450 mL 16 oz 28/400 (450 mL) for PE 8900/Polygen
TFA Deblock Toluene Toluene/TFA Water Content ≤ 200 ppm
L0202502-04 4 x 2.5 L 2.5 L GL45
100/1 TFA Content = 10.3-12.5 g/l
v/w App. of Sol. = clear
DCA Deblock DCM/DCA Water Content ≤ 200 ppm
L020251-04 4 x 2.5 L 2.5 L GL45
97/3 DCA Content = 42.2-51.6 g/l
v/v App. of Sol. = clear
TCA Deblock DCM/TCA Water Content ≤ 200 ppm
L020250-04 4 x 2.5 L 2,5 L GL45
100/3 TCA Content = 28-32 g/l
L020400-04 4 x 4 L 4 L 38/430
v/w App. of Sol. = clear L022500-01 1 x 25 L 25 L Drum, Disposal
L320045-01 1 x 450 mL 16 oz 28/400 (450 mL)
L820090-06 6 x 900 mL 32 oz 28/400 (900 mL) for PE 8900/Polygen
Activator 42 0.1M ACN/Activator42 Water Content ≤ 30ppm
L0302502-04 4 x 2.5 L 2,5 L GL45
100/3 Act. 42 Content = 28-31 g/l
L3300182-06 6 x 180 mL 8 oz 24/400 (200 mL) for ABI
v/w App. of Sol. = clear L8300202-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
L8300452-06 6 x 450 mL 16 oz 28/400 (450 mL)
Table 5. Liquid Reagent Formulations
Solutions for Oligonucleotides SynthesisLiquid Reagents
8
Reagent Description Packaging Available
Formulation Specifications Product Number Unit Bottle Type
Activator 42 0.25 M ACN/Activator42 Water Content ≤ 30 ppm
L0302512-04 4 x 2.5 L 2.5 L GL45
100/7 Act. 42 Content = 70-77 g/l
L8300212-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
v/w App. of Sol. = clear L8300462-06 6 x 450 mL 16 oz 28/400 (450 mL)
ETT Activator 0.25M ACN/ETT Water Content ≤ 30 ppm
L0302501-04 4 x 2.5 L 2.5 L GL45
100/3 ETT Content = 32-35 g/l
L0302001-04 4 x 2 L 2 L 38/430
v/w App. of Sol. = clear L8300201-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
L8300451-06 6 x 450 mL 16 oz 28/400 (450 mL)
DCI Activator 0.25M ACN/DCI Water Content ≤ 30 ppm
L080250-04 4 x 2.5 L 2,5 L GL45
100/3 DCI Content=29-32 g/l L080400-04 4 x 4 L 4 L 38/430
v/w App. of Sol. = clear L082500-01 1 x 25 L 25 L Returnable Drum
L380018-06 6 x 180 mL 8 oz 24/400 (200 mL) for ABI
L880020-06 6 x 200 mL 8 oz 28/400 (200 mL)
L880045-06 6 x 450 mL 16 oz 28/400 (450 mL)
Cap A THF/Ac2O Ac2O Content = 88.7-108.4 g/l
L040250-01 1 x 2.5 L 2.5 L GL45
90.9/9.1 App. of Sol. = clear L040250-04 4 x 2.5 L 2.5 L GL45
v/v L040400-01 1 x 4 L 4 L 38/430
L040400-04 4 x 4 L 4 L 38/430
L040200-04 4 x 2 L 2 L 38/430
L840020-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
L840045-06 6 x 450 mL 16 oz 28/400 (450 mL)
Cap A for ABI instruments
THF/Lutidine/Ac2O Ac2O Content = 97.0-119.0 g/l
L040251-01 1 x 2.5 L 2.5 L GL45
80/10/10 App. of Sol. = clear, colorless to brownish solution
L340018-06 6 x 180 mL 8 oz 24/400 (200 mL)
v/v/v L340045-06 6 x 450 mL 16 oz 28/400 (450 mL)
Cap B THF/Py/NMI Water Content ≤ 200 ppm
L050250-01 1 x 2.5 L 2.5 L GL45
80/10/10 Formulation (GC) conforms
L050250-04 4 x 2.5 L 2.5 L GL45
v/v/v App. of Sol. = clear L050400-01 1 x 4 L 4 L 38/430
L050400-04 4 x 4 L 4 L 38/430
L050200-04 4 x 2 L 2 L 38/430
L850020-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
L850045-06 6 x 450 mL 16 oz 28/400 (450 mL) for PE 8900/Polygen
Table 6. Liquid Reagent Formulations
Solutions for Oligonucleotides SynthesisLiquid Reagents
www.safcsupplysolutions.com
9
Reagent Description Packaging Available
Formulation Specifications Product Number Unit Bottle Type
Cap B for ABI instruments
THF/NMI Water Content ≤ 200 ppm
L050251-01 1 x 2.5 L 2.5 L GL45
84/16 Formulation (GC) conforms
L350018-06 6 x 180 mL 8 oz 24/400 (200 mL)
v/v App. of Sol. = clear L350045-06 6 x 450 mL 16 oz 28/400 (450 mL)
Fast Deprotection Cap A THF/Tac2O Tac2O Content = 45-55 g/l
L070250-01 1 x 2.5 L 2.5 L GL45
100/5 App. of Sol. = clear L070400-01 1 x 4 L 4 L 38/430
v/w L370018-06 6 x 180 mL 8 oz 24/400 (200 mL) for ABI
L370045-06 6 x 450 mL 16 oz 28/400 (450 mL)
L870020-06 6 x 200 mL 8 oz 28/400 (200 mL) PE 8900/Polygen
Oxidizer 0.02M THF/H2O/Py/Iodine Iodine Content = 3.9-4.7 g/l
L060250-04 4 x 2.5 L 2.5 L GL45
90.54/9.05/0.41/0.43 App. of Sol. = brownish L060400-04 4 x 4 L 4 L 38/430
v/v/v/w L060200-04 4 x 2 L 2 L 38/430
L860021-06 6 x 200 mL 8 oz 24/400 (200 mL) for ABI
L860020-06 6 x 200 mL 8 oz 28/400 (200 mL) for PE 8900/Polygen
L860045-06 6 x 450 mL 16 oz 28/400 (450 mL) for PE 8900/Polygen
Oxidizer 0.1M for ABI instruments
THF/H2O/Py/Iodine Iodine Content = 22.9-27.9 g/l
L060251-04 4 x 2.5 L 2.5L GL45
77/2/21/2.54 App. of Sol. = brownish
L360020-06 6 x 200 mL 8 oz 24/400 (200 mL)
v/v/v/w L360045-06 6 x 450 mL 16 oz 28/400 (450 mL)
Oxidizer 0.02M for ABI 3900 instruments
THF/H2O/Py/Iodine Iodine Content = 5.4-6.6 g/l
L060252-04 4 x 2.5 L 2.5 L GL45
66/12/22/0.6 App. of Sol. = brownish
L260045-06 6 x 450 mL 16 oz 28/400 (450 mL)
v/v/v/w
ACN Based Liquid Reagents
Cap A ACN ACN/Ac2O Ac2O Content = 88.7-108.4 g/l
LB40250-01 1 x 2.5 L 2.5 L GL45
90.9/9.1 App. of Sol. = clear
v/v
Cap B ACN ACN/Py/NMI Water Content ≤ 100 ppm
LB50250-01 1 x 2.5 L 2.5 L GL45
80/10/10 Formulation (GC) conforms
v/v/v App. of Sol. = clear
Cap 1 in ACN ACN/Lutidine/NMI Water Content ≤ 200 ppm
LR40250-01 1 x 2.5 L 2.5 L GL45
80/10/10 Formulation (GC) conforms
v/v/v App. of Sol. = clear
Table 7. Liquid Reagent Formulations
Solutions for Oligonucleotides SynthesisLiquid Reagents
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Reagent Description Packaging Available
Formulation Specifications Product Number Unit Bottle Type
Fast Deprotection Cap 2 in ACN
Tac2O/ACN Tac2O Content = 45-55g/l
LR50250-01 1 x 2.5 L 2.5 L GL45
100/5 App. of Sol. = clear
v/w
Oxidizer in ACN ACN/H2O/Py/Iodine Iodine Content = 3.9-4.7 g/l
LR60250-04 4 x 2.5 L 2.5 L GL45
90.54/9.05/0.41/0.43
v/v/v/w
ÄKTA Solution
DCA Deblock (Toluene) DCA/Toluene Water Content ≤ 200 ppm
L520251-04 4 x 2.5 L 2.5 L GL45
97/3 DCA Content = 42.2-51.6
v/v App. of Sol. = clear
BTT Activator 0.3 M 2.5 L, ÄKTA
ACN/BTT Water Content ≤ 30 ppm
L580250-04 4 x 2.5 L 2.5 L GL45
100/6 BTT Content = 51.9-63.4
v/w App. of Sol. = clear
Cap A for Äkta and Oligo Pilot
ACN/NMI Water Content ≤ 100 ppm
L540250-01 1 x 2.5 L 2.5 L GL45
80/20 Formulation (GC) conforms
v/v App. of Sol. = clear
Cap B1 for Äkta and Oligo Pilot
ACN/Ac2O Formulation (GC) conforms
L550250-01 1 x 1.25 L 2.5 L GL45
60/40 App. of Sol. = clear
v/v
Cap B2 for Äkta and Oligo Pilot
ACN/Py Water Content ≤ 200 ppm
L550251-01 1 x 1.25 L 2.5 L GL45
40/60 Formulation (GC) conforms
v/v App. of Sol. = clear
Oxidizer 0,05M for Äkta and Oligo Pilot
Iodine/Py/H2O Iodine Content = 11.4-14.0 g/l
L560250-04 4 x 2.5 L 2.5 L GL45
0,05M/90/10 App. of Sol. = clear
m/v/v
Table 8. Liquid Reagent Formulations
Solutions for Oligonucleotides SynthesisLiquid Reagents
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Quality Control
SAFC has rigorous QC protocol for every product produced to identify purity levels against set specifications, including custom QC testing. State-of-the-art analytical methods include:
• RP & SAX HPLC
• LC-MS
• NMR (31P, 1H, 13C, 19F)
• Nucleic acid synthesis test
• FT-IR
• Titration & “Wet Chemistry”
• UV/VIS
• Karl Fischer water analysis
• Mercury porosimetry
Quality Assurance
As a trusted partner to the oligonucleotide market, SAFC assists its customers in new product development. We know that developing materials includes having insight to their quality requirements. As a result, our company culture fosters continuous process improvement and employs quality systems to provide complete traceability on all our raw materials, processes and procedures. We serve customers conducting audits with complete documentation and files; our quality assurance department routinely accepts requests for documentation, including Certificates of Origin. Our Hamburg facility was one of the first in the industry to be ISO 9001 certified (1993), and we have synthesized amidites and liquid reagents under ISO9000:2000 since then.
Abbreviations
ACN Acetonitrile
DCM Dichloromethane
TCA Trichloroacetic Acid
TFA Trifluoroacetic Acid
DCA Dichloroacetic Acid
THF Tetrahydrofurane
Ac2O Acetic Acid Anhydride
Py Pyridine
NMI N-Methylimidazole
Tac2O tert. Butylphenoxyacetyl Acetic Acid Anhydride
DCI 4,5-Dicyanoimidazole
ETT 5-Ethylthio-1H-tetrazole
KUC04695-5031200049
SAFC®, SAFC Supply Solutions® and Sigma-Aldrich® are registered trademarks of Sigma-Aldrich Biotechnology L.P. and Sigma-Aldrich Co.
Proligo® Reagents is a registered trademark of Sigma-Aldrich Biotechnology L.P. and Sigma-Aldrich Co. Activator® 42 is a registered trademark of Sigma-Aldrich
Biotechnology L.P. and Sigma-Aldrich Co. ABI® is a registered trademark of Applera Corporation or its subsidiaries. ÄKTA™ oligopilot is a trademark of GE
Healthcare. Expedite™ is a trademark of Applera Corporation or its subsidiaries.
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Solutions for Oligonucleotides SynthesisLiquid Reagents
www.safcsupplysolutions.com