PF-543 Citrate_1415562-83-2_DataSheet_MedChemExpress

MagMAX ™ FFPE DNA/RNA Ultra KitAutomated or manual isolation of total nucleic acid (TNA) from FFPE samples using AutoLys tubesCatalog Number A31881Pub. No. MAN0017538 Rev.A.0WARNING! Read the Safety Data Sheets (SDSs) and follow thehandling instructions. Wear appropriate protective eyewear,clothing, and gloves. Safety Data Sheets (SDSs) are available from /support .Product descriptionThe Applied Biosystems ™ MagMAX ™ FFPE DNA/RNA Ultra Kit is designed to isolate both DNA and RNA from the same section offormaldehyde- or paraformaldehyde-fixed, paraffin-embedded (FFPE)tissues. The kit also allows for flexibility to isolate DNA only, RNA only or total nucleic acid (TNA). The kit uses MagMAX ™ magnetic-bead technology, ensuring reproducible recovery of high-quality nucleic acid through manual or automated processing. The isolated nucleic acid is appropriate for use with a broad range of downstream assays, such as quantitative real-time RT-PCR and next-generation sequencing.In addition to the use of traditional solvents, the kit is compatible with Autolys M tubes that enable a faster and more convenient means of deparaffinizing FFPE samples by eliminating the need for organic solvents such as xylene or CitriSolv and ethanol. Samples are put into the tubes for protease digestion, tubes are lifted with the Auto-pliers or Auto-Lifter and then samples are spun down. The wax and debris are contained in the upper chamber while the lysate is passed through.Afterwards, the clarified lysate can be directly purified with the MagMAX ™ FFPE DNA/RNA Ultra Kit.For guides without using AutoLys M tubes for sequential DNA and RNA isolation, or DNA isolation, or RNA isolation only, seeMagMAX ™ FFPE DNA/RNA Ultra Kit User Guide (sequential DNA/RNA isolation) (Pub. No. MAN0015877), or MagMAX ™ FFPE DNA/RNA Ultra Kit User Guide (DNA isolation only) (Pub. No. MAN0015905), or MagMAX ™ FFPE DNA/RNA Ultra Kit User Guide (RNA isolation only)(Pub. No. MAN0015906), respectively.This guide describes isolation of TNA from FFPE tissue blocks or FFPE slides using AutoLys M tubes. Three optimized methods for sections or curls both up to 40 µm using AutoLys M tubes are included:•Manual sample processing.•KingFisher ™ Flex Magnetic Particle Processor with 96 Deep-Well Head (DW96; 96-well deep well setting).•KingFisher ™ Duo Prime Magnetic Particle Processor (12-well deep well setting).For sequential DNA and RNA isolation, or DNA isolation, or RNA isolation only, see MagMAX ™ FFPE DNA/RNA Ultra Kit User Guide (sequential DNA/RNA isolation) (Pub. No. MAN0017541), MagMAX ™FFPE DNA/RNA Ultra Kit User Guide (DNA isolation only)(Pub. No. MAN0017539), or MagMAX ™ FFPE DNA/RNA Ultra Kit User Guide (RNA isolation only) (Pub. No. MAN0017540), respectively.Contents and storageReagents provided in the kit are sufficient for 48 TNA isolations from sections up to 40 µm with the AutoLys workflow.Table 1 MagMAX ™ FFPE DNA/RNA Ultra Kit (Cat. No. A31881)Additional reagents are available separately; Protease Digestion Buffer, Binding Solution, and DNA Wash Buffer are also available as Cat. No. A32796. [2]Shipped at room temperature. [3]Not used in TNA workflow[4]Final volume; see “Isolate TNA“ on page 4.Required materials not suppliedUnless otherwise indicated, all materials are available through . MLS: Fisher Scientific ( ) or other major laboratory supplier.Table 2 Materials required for nucleic acid isolation (all methods)Table 3 Additional materials required for manual isolationTable 4 Additional materials required for automated isolationIf needed, download the KingFisher ™ Duo Prime or Flex programThe programs required for this protocol are not pre-installed on theKingFisher ™Duo Prime Magnetic Particle Processor or on theKingFisher ™Flex Magnetic Particle Processor 96DW.1.On the MagMAX ™ FFPE DNA/RNA Ultra Kit product web page,scroll down to the Product Literature section.2.Right-click on the appropriate program file(s) for your samplesize to download the program to your computer:3.4.Refer to the manufacturer's documentation for instructions forinstalling the program on the instrument.Procedural guidelines•Perform all steps at room temperature (20–25°C) unless otherwise noted.•When mixing samples by pipetting up and down, avoid creating bubbles.•When working with RNA:–Wear clean gloves and a clean lab coat.–Change gloves whenever you suspect that they are contaminated.–Open and close all sample tubes carefully. Avoid splashing or spraying samples.–Use a positive-displacement pipettor and RNase-free pipette tips.–Clean lab benches and equipment periodically with an RNase decontamination solution, such as RNase Zap ™ Solution (Cat.No. AM9780).•Incubation at 60°C can be extended overnight to increase DNA yields, followed by incubation at 90°C for 1 hour.•Volumes for reagent mixes are given per sample. We recommend that you prepare master mixes for larger sample numbers. To calculate volumes for master mixes, refer to the per-well volume and add 5–10% overage.Before you beginBefore first use of the kit•Prepare the Wash Solutions from the concentrates:–Add 46 mL of isopropanol to RNA Wash Buffer Concentrate ,mix, and store at room temperature.–Add 168 mL of ethanol to Wash Solution 2 Concentrate , mix,and store at room temperature.Before each use of the kit•Equilibrate the Nucleic Acid Binding Beads to room temperature.•Pre-heat the incubators or ovens to 60°C and 90°C.•Prepare the following solutions according to the following tables.Table 5 Protease solutionTable 6 TNA Binding BufferPrepare the FFPE samples•For curls from FFPE tissue blocks: proceed to “Prepare the curls from FFPE tissue blocks“ on page 3.•For FFPE slide-mounted sections: proceed to “Prepare samples from FFPE slides“ on page 3.Prepare the curls from FFPE tissue blocksa.Cut sections from FFPE tissue blocks using a microtome.Note: For miRNA extraction, we recommend using sections of 10 µm or thicker.b.Collect each section in an AutoLys M tube.1Section FFPE tissue blocks a.Add 235 µL of the Protease Solution (see Table 5).Note: If working with curls, they might stick straight up so make sure to submerge samples in theProtease Solution with a tip or a 1 mL syringe plunger or do a quick spin down at 3000 rpm for 1 minute prior to the addition of buffer to collapse the curl. Time may be extended.b.Incubate at 60°C for 1 hour or longer.Note: Use the AutoLys racks and place in an incubator or oven.c.Incubate at 90°C for 1 hour.Note: For automated isolation, set up the processing plates during the incubation.·For isolation using KingFisher ™ Duo Prime Magnetic Particle Processor, proceed to “Set up the processing plate“ on page 4.·For isolation using KingFisher ™ Flex Magnetic Particle Processor 96DW, proceed to “Set up the TNA processing plates“ on page 5.2Digest with Protease a.Allow samples to cool down for 3–5 minutes before proceeding to lift the tubes.e the Auto-plier for individual tube lifting or the Auto-lifter for multiple tube lifting of up to 24 tubes.c.Lock the tubes in position by hand or use the locking lid.d.Centrifuge at 2000 × g for 10 minutes in a benchtop centrifuge with plate adapters.e.Unlock the tubes by hand or remove the locking lid.e the Auto-plier or Auto-lifer to lift the inner tube for sample access.g.Proceed to purification. See “Isolate TNA“ on page 43Lift the tubesPrepare samples from FFPE slidesa.Pipet 2–4 µL of Protease Digestion Buffer depending on the tissue size evenly across the FFPE tissuesection on the slide to pre-wet the section.Note: You can adjust the volume of Protease Digestion Buffer if the tissue is smaller or larger.b.Scrape the tissue sections in a single direction with a clean razor blade or scalpel, then collect the tissueon the slide into a cohesive mass.c.Transfer the tissue mass into an AutoLys M tube with the scalpel or a pipette tip.d.Add 235 µL of the Protease Solution (see Table 5).Note: Be sure to submerge samples in the Protease Solution with a tip or a 1 mL syringe plunger e.Incubate at 60°C for 1 hour or longer.Note: Use the AutoLys racks and place in an incubator or oven.f.Incubate at 90°C for 1 hour.Note: For automated isolation, set up the processing plates during the incubation.·For isolation using KingFisher ™ Duo Prime Magnetic Particle Processor, proceed to “Set up the processing plate“ on page 4.·For isolation using KingFisher ™ Flex Magnetic Particle Processor 96DW, proceed to “Set up the TNA processing plates“ on page 5.4Scrape the samples and digest with Proteasea.Allow samples to cool down for 3–5 minutes before proceeding to lift the tubes.e the Auto-plier for individual tube lifting or the Auto-lifter for multiple tube lifting of up to 24 tubes.c.Lock the tubes in position by hand or use the locking lid.d.Centrifuge at 2000 × g for 10 minutes in a benchtop centrifuge with plate adapters.e.Unlock the tubes by hand or remove the locking lid.e the Auto-plier or Auto-lifer to lift the inner tube for sample access.g.Proceed to purification. See “Isolate TNA“ on page 45Lift the tubesIsolate TNA•To isolate TNA manually, proceed to “Isolate TNA manually“ on page 4.•To isolate TNA using the KingFisher ™Duo Prime Magnetic Particle Processor, proceed to “Isolate TNA using KingFisher ™ Duo Prime Magnetic Particle Processor“ on page 4.•To isolate TNA using the KingFisher ™Flex Magnetic Particle Processor 96DW, proceed to “Isolate TNA using KingFisher ™ Flex Magnetic Particle Processor 96DW“ on page 5.Isolate TNA manuallyUse microcentrifuge tubes to perform manual TNA isolations.a.After the Protease digestion is complete, add 20 µL of Nucleic Acid Binding Beads to the samples.b.Add 900 µL of TNA Binding Buffer (see Table 6) to the sample.c.Shake for 5 minutes at speed 10 or 1150 rpm.d.Place the sample on the magnetic stand for 2 minutes or until the solution clears and the beads are pelleted against the magnet.e.Carefully discard the supernatant with a pipette.1Bind the TNA to beadsa.Wash the beads with 500 µL of RNA Wash Buffer.b.Shake for 1 minute at speed 10 or 1150 rpm until the mixture is thoroughly chocolate brown in color.c.Place the sample on the magnetic stand for 2 minutes or until the solution clears and the beads arepelleted against the magnet.d.Carefully discard the supernatant with a pipette.e.Repeat steps a-d.f.Wash the beads with 500 µL of Wash Solution 2.g.Shake for 1 minute at speed 10 or 1150 rpm until the mixture is thoroughly chocolate brown in color.h.Place the sample on the magnetic stand for 2 minutes or until the solution clears and the beads arepelleted against the magnet.i.Carefully discard the supernatant with a pipette.j.Repeat steps f-i.k.Shake for 1–2 minutes at speed 10 or 1150 rpm to dry the beads.Do not over-dry the beads. Over-dried beads results in low TNA recovery yields.2Wash TNA on the beadsa.Add 50 µL of Elution Solution to the beads.b.Shake for 5 minutes at speed 10 or 1150 rpm and at 55°C until the mixture is thoroughly chocolatebrown in color.c.Place the sample on the magnetic stand for 2 minutes or until the solution clears and the beads arepelleted against the magnet.The supernatant contains the purified TNAThe purified TNA is ready for immediate use. Store at –20°C or –80°C for long-term storage.3Elute the TNAIsolate TNA using KingFisher ™ Duo Prime Magnetic Particle ProcessorDuring the protease incubation, add processing reagents to the wells of a MagMAX ™ Express-96 Deep WellPlate as indicated in the following table.Table 7 TNA plate setupRow on the MagMAX ™ Express-96 Deep Well Plate.4Set up the processing plate a.Ensure that the instrument is set up for processing with the deep well 96–well plates and select theappropriate program A31881_DUO_large_vol_TNA on the instrument.b.At the end of the protease incubation, add 200 µL of sample to each well in Row G of the TNA plate.c.Add 20 µL of Nucleic Acid Binding Beads to each sample well in Row G.d.Start the run and load the prepared processing plates when prompted by the instrument.5Bind, wash, rebind, and elute the TNA5Bind, wash, rebind,and elute the TNA(continued)e.At the end of the run, remove the Elution Plate from the instrument and transfer the eluted TNA(Row A of TNA plate) to a new plate and seal immediately with a new MicroAmp ™ Clear Adhesive Film.IMPORTANT! Do not allow the purified samples to sit uncovered at room temperature for more than 10minutes, to prevent evaporation and contamination.The purified TNA is ready for immediate use. Store at –20°C or –80°C for long-term storage.Isolate TNA using KingFisher ™ Flex Magnetic Particle Processor 96DWDuring the protease incubation, add processing reagents to the wells of MagMAX ™ Express-96 Plates asindicated in the following table.Table 8 TNA plates setupPosition on the instrument6Set up the TNA processing platesa.Ensure that the instrument is set up for processing with the deep well magnetic head and select theA31881_FLEX_large_vol_TNA program on the instrument.b.At the end of the protease incubation, add 200 µL of sample to each well in Plate 1.c.Add 20 µL of Nucleic Acid Binding Beads to each sample well in Plate 1.d.Start the run and load the prepared processing plates in their positions when prompted by theinstrument (see “Set up the TNA processing plates“ on page 5).e.At the end of the run, remove the Elution Plate from the instrument and seal immediately with a newMicroAmp ™ Clear Adhesive Film.IMPORTANT! Do not allow the purified samples to sit uncovered at room temperature for more than 10minutes, to prevent evaporation and contamination.The purified TNA is ready for immediate use. Store at –20°C or –80°C for long-term storage. .7Bind, wash, rebind, and elute the TNALimited product warrantyLife Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and Conditions of Sale found on Life Technologies' website at /us/en/home/global/terms-and-conditions.html . If you have any questions,please contact Life Technologies at /support .Manufacturer: Life Technologies Corporation | 2130 Woodward Street | Austin, TX 78744The information in this guide is subject to change without notice.DISCLAIMER : TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE,MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.Important Licensing Information : This product may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all applicable Limited Use Label Licenses.©2018 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified./support | /askaquestion 。

非药用类麻醉药品和精神药品管制品种目录（2015年版）
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中文名 N-(2-甲氧基苄基)-2-(2,5-二甲氧基-4-溴苯基)乙胺 2,5-二甲氧基-4-氯苯乙胺 N-(2-甲氧基苄基)-2-(2,5-二甲氧基-4-氯苯基)乙胺 2,5-二甲氧基-4-甲基苯乙胺 N-(2-甲氧基苄基)-2-(2,5-二甲氧基-4-甲基苯基)乙胺 2,5-二甲氧基-4-乙基苯乙胺 N-(2-甲氧基苄基)-2-(2,5-二甲氧基-4-碘苯基)乙胺 2,5-二甲氧基-4-丙基苯乙胺 2,5-二甲氧基-4-乙硫基苯乙胺 2,5-二甲氧基-4-异丙基硫基苯乙胺 2,5-二甲氧基-4-丙硫基苯乙胺
12 2-氟苯丙胺 13 2-氟甲基苯丙胺 14 1-(2-苯并呋喃基)-N-甲基-2-丙胺 15 3-氟苯丙胺 16 3-氟甲基苯丙胺 17 4-氯苯丙胺 18 4-氟苯丙胺 19 4-氟甲基苯丙胺 20 1-[5-(2,3-二氢苯并呋喃基)]-2-丙胺 21 1-(5-苯并呋喃基)-N-甲基-2-丙胺 22 6-溴-3,4-亚甲二氧基甲基苯丙胺 23 6-氯-3,4-亚甲二氧基甲基苯丙胺 24 1-(2,5-二甲氧基-4-氯苯基)-2-丙胺 25 1-(2-噻吩基)-N-甲基-2-丙胺 26 N-(1-氨甲酰基-2-甲基丙基)-1-(5-氟戊基)吲哚-3-甲酰胺 27 N-(1-氨甲酰基-2-甲基丙基)-1-(5-氟戊基)吲唑-3-甲酰胺 28 N-(1-氨甲酰基-2,2-二甲基丙基)-1-(5-氟戊基)吲哚-3-甲酰胺 29 N-(1-甲氧基羰基-2-甲基丙基)-1-(5-氟戊基)吲唑-3-甲酰胺 30 N-(1-金刚烷基)-1-(5-氟戊基)吲唑-3-甲酰胺 31 1-(5-氟戊基)吲哚-3-甲酸-8-喹啉酯 32 1-(5-氟戊基)-3-(2,2,3,3-四甲基环丙甲酰基)吲哚 33 1-[2-(N-吗啉基)乙基]-3-(2,2,3,3-四甲基环丙甲酰基)吲哚 34 1-(4-四氢吡喃基甲基)-3-(2,2,3,3-四甲基环丙甲酰基)吲哚 35 N-(1-氨甲酰基-2-甲基丙基)-1-(环己基甲基)吲唑-3-甲酰胺 36 N-(1-氨甲酰基-2-甲基丙基)-1-(4-氟苄基)吲唑-3-甲酰胺 37 N-(1-氨甲酰基-2-甲基丙基)-1-戊基吲唑-3-甲酰胺 38 N-(1-氨甲酰基-2,2-二甲基丙基)-1-戊基吲哚-3-甲酰胺 39 N-(1-氨甲酰基-2,2-二甲基丙基)-1-戊基吲唑-3-甲酰胺 40 1-[(N-甲基-2-哌啶基)甲基]-3-(1-萘甲酰基)吲哚 41 1-[(N-甲基-2-哌啶基)甲基]-3-(1-金刚烷基甲酰基)吲哚 42 1-[(N-甲基-2-哌啶基)甲基]-3-(2-碘苯甲酰基)吲哚 43 N-(1-金刚烷基)-1-戊基吲哚-3-甲酰胺 44 N-(1-金刚烷基)-1-戊基吲唑-3-甲酰胺 45 1-(1-萘甲酰基)-4-戊氧基萘 46 N-(1-甲基-1-苯基乙基)-1-(4-四氢吡喃基甲基)吲唑-3-甲酰胺 47 1-(5-氟戊基)-3-(4-乙基-1-萘甲酰基)吲哚 48 1-(4-氟苄基)-3-(1-萘甲酰基)吲哚 49 1-(4-氟苄基)吲哚-3-甲酸-8-喹啉酯 50 2-甲基-1-戊基-3-(1-萘甲酰基)吲哚 51 2-甲基-1-丙基-3-(1-萘甲酰基)吲哚 52 1-己基-3-(1-萘甲酰基)吲哚 53 1-戊基-3-(4-甲氧基-1-萘甲酰基)吲哚 54 1-戊基-3-(4-甲基-1-萘甲酰基)吲哚 55 1-戊基-3-(2-氯苯乙酰基)吲哚 56 1-戊基-3-(4-乙基-1-萘甲酰基)吲哚 57 1-戊基-2-(2-甲基苯基)-4-(1-萘甲酰基)吡咯 58 1-(5-氟戊基)-3-(4-甲基-1-萘甲酰基)吲哚 59 N-(1-甲氧基羰基-2,2-二甲基丙基)-1-(环己基甲基)吲哚-3-甲酰胺 60 N-(1-甲氧基羰基-2,2-二甲基丙基)-1-(4-氟苄基)吲唑-3-甲酰胺 61 1-戊基吲哚-3-甲酸-8-喹啉酯 62 N-(1-氨甲酰基-2-苯基乙基)-1-(5-氟戊基)吲唑-3-甲酰胺

MAC4LDFpis Rev 04/221Product InformationMetaPolyzyme, DNA freeSuitable for Microbiome researchMAC4LDFSynonym: Multilytic Enzyme Mix Storage Temperature –20 °CProduct DescriptionMetagenomics investigates all DNA that has been isolated directly from given single samples, such as environmental samples or biological organisms.1,2Metagenomics allows for the investigation of microbes that exist in extreme environments, and which have been historically difficult to isolate, culture, andstudy.3 Metagenomics has revealed the existence of novel microbial species.4 Applications ofmetagenomics work include public health dataanalysis,5,6 discovery of novel proteins, enzymes and natural products,7,8 environmental studies,9,10 and agricultural investigations.11,12Microbes are difficult to disrupt because the cell walls may form capsules or resistant spores. DNA can be extracted by using lysing enzymes such as lyticase, chitinase, zymolase, and gluculase to induce partial spheroplast formation. Spheroplasts are subsequently lysed to release DNA.MetaPolyzyme products (Cat. Nos. MAC4L, MAC4LDF) are based on a multi-lytic enzyme mixture, originally developed by Scott Tighe, for use in microbiome and DNA extraction efficiency studies, and formulated for effective lysis of microbiome samples from extreme environments. MetaPolyzyme was originally evaluated and developed in consultation and collaboration with the Association of Biomolecular Resource Facilities (ABRF) Metagenomics and Microbiome ResearchGroup (MMRG; formerly the Metagenomics Research Group, MGRG).13-16Studies of microbial communities have beenenhanced by the use of culture-independent analytical techniques such as 16S rRNA gene sequencing and metagenomics. DNA contamination during sample preparation is a major problem of sequence-based approaches. Extraction reagents free of DNA contaminants are thus essential. MetaPolyzyme, DNA free was developed to address the need for DNA-free reagents, to minimize microbial DNA contamination from reagents. This productundergoes strict quality control testing to ensure the absence of detectable levels of contaminatingmicrobial DNA using 35 cycles PCR amplification of 16S and 18S rDNA using universal primer sets.Precautions and DisclaimerFor R&D use only. Not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.ReagentThe enzymes in MetaPolyzyme, DNA free are:• Mutanolysin • Achromopeptidase • Lyticase • Chitinase • Lysostaphin •LysozymeAll the enzymes are individually tested for theabsence of contaminating DNA using 16S and 18S PCR amplification.Mutanolysin (from Streptomyces globisporus )Mutanolysin is a muralytic enzyme (muramidase) that cleaves the β-N -acetylmuramyl-(1→4)-N -acetylglucosamine linkage of the bacterial cell wall peptidoglycan-polysaccharide, particularly the β(1→4) bond in MurNAc-GlcNAc.17 Mutanolysin particularly acts on many Gram-positive bacteria, where the enzyme’s carboxy -terminal moietiesparticipate in the recognition and binding of unique cell wall structures.MAC4LDFpis Rev 04/22AchromopeptidaseAchromopeptidase (known also as β-lytic protease 18) has potent bacteriolytic activity on many Gram-positive aerobic bacteria 19 with high lytic activity, against bacterial strains with the A1α chemotype (such as Aerococcus viridans ), and the A3αchemotype (such as Staphylococcus epidermidis ) for cell wall peptidoglycan structures. The enzyme has been reported to have particular recognition for Gly-X sites in peptide sequences, and for Gly-Gly and ᴅ-Ala-X sites in peptidoglycans.20Lyticase (from Arthrobacter luteus )Lyticase is useful in digestion of linear glucosepolymers with β(1→3) linkages, of yeast glycan coats and for spheroplast formation, and of the cell wall of active yeast cells.Chitinase (from Streptomyces griseus )Chitinase degrades chitin by enzymatic hydrolysis to N-acetyl-D-glucosamine. Degradation occurs via two consecutive enzyme reactions: •Chitodextrinase-chitinase, apoly(1,4-β-[2-acetamido-2-deoxy-D-glucoside])-glycanohydrolase, removes chitobiose units from chitin.•N-acetylglucosaminidase-chitobiase cleaves the disaccharide to its monomer subunits, N-acetyl-D-glucosamine (NAGA).Lysostaphin (from Staphylococcus staphylolyticus )Lysostaphin is a lytic enzyme with activity against Staphylococcus species, including S. aureus . Lysostaphin has hexosaminidase, amidase, and endopeptidase activities. It cleaves polyglycine crosslinks in the cellular wall of Staphylococcus species, which leads to cell lysis.21,22Lysozyme (from chicken egg white)Lysozyme hydrolyzes β(1→4) linkages betweenN -acetylmuraminic acid and N -acetyl-D-glucosamine residues in peptidoglycan, and betweenN -acetyl-D-glucosamine residues in chitodextrin. Lysozyme lyses the peptidoglycan cell wall of Gram-positive bacteria.23Storage/StabilityThis product ships at cooler temperature conditions. Long-term storage at –20 °C is recommended. Reconstituted solutions of MetaPolyzyme, DNA free may be stored at –20 °C, but long-term solution stability has not been examined.Preparation InstructionsBecause of the great diversity of samples formetagenomics studies, it will be necessary for each researcher to work out particular conditions for optimal sample preparation and treatment. It is recommended to reconstitute MetaPolyzyme, DNA free in sterile PBS buffer, pH 7.5 (no EDTA, calcium or magnesium present in solution). The following is a sample procedure, to be scaled appropriately:1. Add 100 µL of sterile PBS (pH 7.5) to 1 vial ofMetaPolyzyme, DNA free.1.1. Resuspend by gentle agitation or pipetting. 1.2. Set solution aside at 2-8 °C until Step 7. 2. Thoroughly suspend sample in sterile PBS(pH 7.5). 3. Add 200 µL of sample in PBS to a 2 mLpolypropylene microcentrifuge tube. 4. Optional pellet wash:4.1. To sample tube, add 1 mL of PBS (pH 7.5). 4.2. Vortex, centrifuge and remove supernatant. 4.3. Repeat pellet wash two more times ifneeded. 5. Resuspend pelleted sample in 150 µL of PBS(pH 7.5). Vortex thoroughly.6. Optional: if solution will sit for more than 4 hours,sodium azide may be added to 0.02%. 7. Add 20 µL (*) of MetaPolyzyme, DNA free tosample solution. 8. Incubate at 35 °C for 2-24 hours.9. Continue with standard DNA extraction protocol. (*) The optimal volume and concentration of MetaPolyzyme, DNA free may vary in different experiments.References1. Gilbert, J.A., and Dupont, C.L., Ann. Rev. MarineSci., 3, 347-371 (2011). 2. Kang, H.S., and Brady, S.F., J. Am. Chem. Soc.,136(52), 18111-18119 (2014). 3. Ufarté, L. et al., Biotechnol. Adv., 33(8),1845-1854 (2015). 4. Davison, M. et al., Photosynth. Res., 126(1),135-146 (2015). 5. Afshinnekoo, E. et al., Cell Syst., 1(1), 72-87(2015).The life science business of Merck operatesas MilliporeSigma in the U.S. and Canada.Merck and Sigma-Aldrich are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates.All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources.© 2022 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.MAC4LDFpis Rev 04/22 DK,DT,GCY,TJ,RBG,SBC,MAM36.The MetaSUB International Consortium,Microbiome, 4, 24 (2016). [Erratum inMicrobiome, 4, 45 (2016).]7.Trinidade, M. et al., Front. Microbiol., 6, 890(2015).8.Coughlan, L.M. et al., Front. Microbiol., 6, 672(2015).9.Palomo, A. et al., ISME J., 10(11), 2569-2581(2016).10.Pold, G. et al., Appl. Environ. Microbiol., 82(22),6518-6530 (2016).11.Mitra, N. et al., J. Gen. Virol., 97(8), 1771-1784(2016).12.Theuns, S. et al., Infect. Genet. Evol., 43,135-145 (2016).13.Baldwin, D.A. et al., "Life at the Extreme", ABRFMetagenomics Research Group Poster 2015,presented at the ABRF 2015 Conference, St.Louis, MO, USA, March 28-31, 2015.14.Baldwin, D.A. et al., "Implementing NewStandards in Metagenomics and the ExtremeMicrobiome Project", ABRF MetagenomicsResearch Group Poster 2016, presented at theABRF 2016 Conference, Fort Lauderdale, FL, USA, February 20-23, 2016.15.McIntyre, A. et al., "Life at the Extreme: TheABRF Metagenomics Research Group", ABRFMetagenomics Research Group Poster 2017,presented at the ABRF 2017 Conference, SanDiego, CA, March 25-28, 2017.16.Tighe, S. et al., J. Biomol. Tech., 28(1), 31-39(2017).17.Gründling, A., and Schneewind, O., J. Bacteriol.,188(7), 2463-2472 (2006).18.Li, S.L. et al., J. Bacteriol., 172(11), 6506-6511(1990).19.Ezaki, T., and Suzuki, S., J. Clin. Microbiol.,16(5), 844-846 (1982). 20.Li, S. et al., J. Biochem., 124(2), 332-339(1998).21.Browder, H.P. et al., Biochem. Biophys. 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Product Data SheetPerkadox 14-40K-PD Di(tert-butylperoxyisopropyl) benzenePerkadox® 14-40K-PD is a 40% formulation on an inert carrier system (main carrier clay) in powder form.CAS number2212-81-9, 25155-25-3EINECS/ELINCS No.218-664-7TSCA statuslisted on inventoryMolecular weight338.5Active oxygen contentperoxide9.45%Concentration3.7-3.9%SpecificationsAppearance Off-white powderAssay39.0-41.0 %ApplicationsPerkadox® 14-40K-PD is a bifunctional peroxide which is used for the crosslinking of natural rubber and synthetic rubbers, as well as polyolefins. Rubber compounds containing Perkadox® 14-40K-PD have excellent scorch safety, and under certain conditions one step mixing is possible. Safe processing temperature: 135°C (rheometer ts2 > 20 min.). Typical crosslinking temperature: 175°C (rheometer t90 about 12 min.).Thermal stabilityOrganic peroxides are thermally unstable substances, which may undergo self-accelerating decomposition. The lowest temperature at which self-accelerating decomposition of a substance in the original packaging may occur is the Self-Accelerating Decomposition Temperature (SADT). The SADT is determined on the basis of the Heat Accumulation Storage Test.SADT80°CMethod The Heat Accumulation Storage Test is a recognized test method for thedetermination of the SADT of organic peroxides (see Recommendations on theTransport of Dangerous Goods, Manual of Tests and Criteria - United Nations, NewYork and Geneva).StorageDue to the relatively unstable nature of organic peroxides a loss of quality can be detected over a period of time. To minimize the loss of quality, Nouryon recommends a maximum storage temperature (Ts max. ) for each organic peroxide product.Ts Max.30°CNote When stored under the recommended storage conditions, Perkadox® 14-40K-PDwill remain within the Nouryon specifications for a period of at least 6 months afterdelivery.Packaging and transportThe standard packaging is a cardboard box for 50 lbs peroxide formulation. Both packaging and transport meet the international regulations. For the availability of other packed quantities consult your Nouryon representative. Perkadox®14-40K-PD is classified as Flammable solid, class 4. 1, UN 1325.Safety and handlingKeep containers tightly closed. Store and handle Perkadox® 14-40K-PD in a dry well-ventilated place away from sources of heat or ignition and direct sunlight. Never weigh out in the storage room. Avoid contact with reducing agents (e. g. amines), acids, alkalines and heavy metal compounds (e. g. accelerators, driers and metal soaps). Please refer to the Safety Data Sheet (SDS) for further information on the safe storage, use and handling of Perkadox® 14-40K-PD. This information should be thoroughly reviewed prior to acceptance of this product. The SDS is available at /sds-search.Major decomposition productsMethane, Acetone, tert-Butanol, Di(2-hydroxyisopropyl)benzene, Diacetylbenzene, Acetyl 2-hydroxyisopropyl benzeneAll information concerning this product and/or suggestions for handling and use contained herein are offered in good faith and are believed to be reliable.Nouryon, however, makes no warranty as to accuracy and/or sufficiency of such information and/or suggestions, as to the product's merchantability or fitness for any particular purpose, or that any suggested use will not infringe any patent. Nouryon does not accept any liability whatsoever arising out of the use of or reliance on this information, or out of the use or the performance of the product. Nothing contained herein shall be construed as granting or extending any license under any patent. Customer must determine for himself, by preliminary tests or otherwise, the suitability of this product for his purposes.The information contained herein supersedes all previously issued information on the subject matter covered. The customer may forward, distribute, and/or photocopy this document only if unaltered and complete, including all of its headers and footers, and should refrain from any unauthorized use. Don’t copythis document to a website.Perkadox® is a registered trademark of Nouryon Functional Chemicals B. V. or affiliates in one or more territories.Contact UsPolymer Specialties Americas************************Polymer Specialties Europe, Middle East, India and Africa*************************Polymer Specialties Asia Pacific************************2022-6-30© 2022Polymer crosslinking Perkadox 14-40K-PD。