Shop All Modifying Enzymes

S1 Nuclease (100 U/µL) Thermo Scientific™

Thermo Scientific S1 Nuclease degrades single-stranded nucleic acids, releasing 5'-phosphoryl mono- or oligonucleotides. It is five times more active on DNA than on RNA. S1 Nuclease also cleaves dsDNA at the single-stranded region caused by a nick, gap, mismatch or loop. S1 Nuclease exhibits 3'-phosphomonoesterase activity.

The enzyme is a glycoprotein with carbohydrate content of 18%.

Applications

• Removal of single-stranded overhangs of DNA fragments
• S1 transcript mapping
• Cleavage of hairpin loops
• Creation of unidirectional deletions in DNA fragments in conjunction with Exonuclease III

Note

S1 Nuclease can introduce breaks into double-stranded DNA, RNA and DNA/RNA hybrids at high enzyme and low salt concentrations.

Uracil-DNA Glycosylase (1 U/µL) Thermo Scientific™

Thermo Scientific Uracil-DNA Glycosylase (UDG, UNG) catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA (see Figure 1 in Supporting Data). The enzyme shows no activity on RNA.

Highlights

Active in Thermo Scientific buffers for restriction enzymes and thermophilic polymerases

Applications

• Control of carry-over contamination in PCR
• Glycosylase mediated single nucleotide polymorphism detection (GMPD)
• Site-directed mutagenesis
• As a probe for protein-DNA interaction studies
• SNP genotyping
• Cloning of PCR products
• Generation of single strand overhangs of PCR products and cDNA

Note

The abasic sites formed in DNA by Uracil-DNA Glycosylase may be subsequently cleaved by heat, alkali-treatment or endonucleases that cleave specifically at abasic sites. UDG (UNG) is active in the presence or absence of divalent cations.

Use of this enzyme in certain applications may be covered by patents and may require a license.

RapidOut DNA Removal Kit Thermo Scientific™

Thermo Scientific RapidOut DNA Removal Kit rapidly and safely removes genomic DNA from total RNA and mRNA preparations. Complete digestion of DNA and safe removal of DNase I from the digestion reaction is ensured without RNA damaging steps, such as heating or organic extraction. First, the RNA sample is treated with recombinant RNase-free DNase I to levels below the limit of detection by routine PCR. DNase I is safely removed subsequently using proprietary DNase Removal Reagent (DRR).

DRR efficiently binds DNase I and the complex is collected at the bottom of the tube by centrifugation. The purified RNA is collected as a supernatant. The RNA after RapidOut procedure is free from DNA contamination and free of DNase I. It is ready to use in different applications including end-point or real-time RT-PCR, cloning, microarrays, and Northern blotting.

Highlights

Efficient—complete ds and ssDNA digestion and proprietary technology for DNase I removal
Rapid—single step sufficient for complete DNase I removal
Safe—no need for toxic organic extractions or RNA-damaging heating steps

Applications

Main Applications
• RNA isolation and RNA analysis, particularly RT-qPCR and RT-PCR (customers performing expression analysis of low transcription level genes.
• Customers performing ds-cDNA synthesis from total RNA preps.

Other applications
• Elimination of DNA from RNA for microinjections and transfection.
• Elimination of DNA from RNA prior microarray analysis.
• Elimination of DNA from RNA prior Northern blot analysis.

TrueCut™ Cas9 Protein v2 Invitrogen™

Invitrogen TrueCut Cas9 Protein v2 is a next-generation CRISPR Cas9 protein engineered to deliver maximum editing efficiency. Features include:

• Consistently high editing efficiency in all tested cell lines including standard, immune, primary, and stem, with up to 2X higher editing efficiency in difficult targets than the competition
• High quality—manufactured under strict ISO 13485 quality standards
• Validated protocols for a large number of cell types help you achieve success faster

TrueCut Cas9 Protein v2 is recombinant Streptococcus pyogenes Cas9 (wt) protein, purified from E. coli, for genome editing with CRISPR technology. Cas9 protein forms a very stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9 system. Incorporation of nuclear localization signals (NLS) aids delivery to the nucleus, increasing the rate of genomic DNA cleavage.

• Transfection-ready, using either lipid-mediated transfection reagents or electroporation
• Eliminates time-consuming cloning steps
• Available in 2 concentrations:
    --1 μg/μL for use in standard editing scenarios
    --5 μg/μL for optimization of editing conditions in more difficult scenarios such as in primary or embryonic cells, microinjection, or when screening multiple gRNA sequences at a time
    --For custom sizes or concentrations, please inquire

Options for obtaining CRISPR gRNA for use with TrueCut Cas9 Protein v2:
1. Order transfection-ready TrueGuide synthetic gRNAs
2. Generate transfection-ready gRNA in as little as four hours including template assembly with our GeneArt Precision gRNA Synthesis Kit
3. Have our Custom Services team design, synthesize, and purify in vitro (IVT) gRNA sequences for you

Pyrophosphatase, inorganic (0.1 U/µL) Thermo Scientific™

Thermo Scientific Pyrophosphatase, Inorganic catalyzes the hydrolysis of inorganic pyrophosphate into two orthophosphates. The enzyme requires a divalent metal cation with Mg2+ conferring the highest activity.

Highlights

• Active in Thermo Scientific buffers for DNA polymerases, RNA polymerases, and Reverse Transcriptases

Applications

• High yield RNA synthesis by in vitro transcription (see Reference 2)
• DNA polymerization reactions: preventing accumulation of pyrophosphate (see References 3, 4)
• Removal of contaminant PPi in reagents used for SNP genotyping by methods based on the detection of pyrophosphate

Note

The enzyme can be diluted with the supplied storage (dilution) buffer. Use of this enzyme in certain applications may be covered by patents and may require a license.

RiboLock RNase Inhibitor (40 U/µL) Thermo Scientific™

Thermo Scientific RiboLock RNase Inhibitor inhibits the activity of RNases A,B and C by binding them in a noncompetitive mode at a 1:1 ratio. It does not inhibit eukaryotic RNases T1, T2, U1, U2, CL3 as well as prokaryotic RNases I and H.

Highlights

Performs under a wide range of reaction conditions
Protects RNA from degradation at temperatures up to 55°C

Note

DTT provided in the Storage Buffer ensures stability during long term storage, but is not necessary for inhibitor activity. Recommended concentration 1 U/ µL of a reaction mixture.

DNase I, RNase-free (1 U/µL) Thermo Scientific™

Thermo Scientific DNase I, RNase-free is an endonuclease that digests single- and double-stranded DNA. It hydrolyzes phosphodiester bonds producing mono- and oligodeoxyribonucleotides with 5'-phosphate and 3'-OH groups.

The enzyme activity is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions.

In the presence of Mg2+, DNase I cleaves each strand of dsDNA independently in a statistically random fashion. In the presence of Mn2+, the enzyme cleaves both DNA strands at approximately the same site, producing DNA fragments with blunt-ends or with overhang termini of only one or two nucleotides.

Highlights

• Recombinant enzyme
• Purified from non-animal host with a lower level of intrinsic RNases

Applications

• Preparation of DNA-free RNA
• Removal of template DNA following in vitro transcription
• Preparation of DNA-free RNA prior to RT-PCR and RT-qPCR
• DNA labeling by nick-translation in conjunction with DNA Polymerase I
• Studies of DNA-protein interactions by DNase I, RNase-free footprinting
• Generation of a library of randomly overlapping DNA inserts. Reaction buffer containing Mn2+ is used

Note

DNase I is sensitive to physical denaturation. Mix gently by inverting the tube. Do not vortex.

FastAP Thermosensitive Alkaline Phosphatase (1 U/µL) Thermo Scientific™

Thermo Scientific FastAP Thermosensitive Alkaline Phosphatase catalyzes the release of 5'- and 3'-phosphate groups from DNA, RNA, and nucleotides. This enzyme also removes phosphate groups from proteins.

FastAP is a novel alkaline phosphatase, which is active in all Thermo Scientific restriction enzyme buffers as well as in PCR buffers. It dephosphorylates all types of DNA ends (blunt, 5'- and 3'-overhangs) in 10 minutes at 37°C. The enzyme is inactivated in 5 minutes at 75°C (see Figure 1 in Supporting Data). Therefore, removal of alkaline phosphatase is not required prior to ligation.

Highlights

Recombinant enzyme
Fast dephosphorylation—10 minutes at 37°C
Fast and complete inactivation—5 minutes at 75°C
Simultaneous digestion and dephosphorylation of vector DNA
100% active in restriction enzyme and PCR buffers
PCR clean-up in conjunction with Exo I
Protein dephosphorylation

One protocol for all types of DNA ends:
• 5'-overhangs
• 3'-overhangs
• blunt-ends
• single nucleotides

Applications

• Dephosphorylation of cloning vector DNA to prevent recircularization during ligation
• Simultaneous digestion and dephosphorylation of vector DNA
• PCR product clean-up: nucleotide degradation prior to sequencing of PCR product
• Dephosphorylation of nucleic acid 5'-termini prior to labeling with T4 Polynucleotide Kinase
• Other applications where dephosphorylation of DNA and RNA substrates is necessary
• Protein dephosphorylation

Note

• Binding of FastAP Thermosensitive Alkaline Phosphatase to DNA may result in a band shift in agarose gels. To avoid this, incubate samples with 6X DNA Loading Dye & SDS Solution at 65°C for 10 minutes, and chill on ice prior to electrophoresis.
• FastAP Thermosensitive Alkaline Phosphatase is active in all restriction enzyme buffers and may be added directly to digested DNA. Heat inactivation of the restriction enzyme before dephosphorylation reaction is not necessary.

Agarase (0.5 U/µL) Thermo Scientific™

Thermo Scientific Agarase is an enzyme that specifically digests the agarose polysaccharide core into neoagaro-oligosaccharides. Agarase allows for gentle yet efficient recovery of DNA or RNA fragments from low melting point agarose. The recovered nucleic acids can be directly used for amplification, cloning, sequencing, etc.

DNase I, RNase-free, HC (50 U/µL) Thermo Scientific™

Thermo Scientific DNase I, RNase-free is an endonuclease that digests single- and double-stranded DNA. It hydrolyzes phosphodiester bonds producing mono- and oligodeoxyribonucleotides with 5'-phosphate and 3'-OH groups.

The enzyme activity is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions.

In the presence of Mg2+, DNase I cleaves each strand of dsDNA independently in a statistically random fashion. In the presence of Mn2+, the enzyme cleaves both DNA strands at approximately the same site, producing DNA fragments with blunt-ends or with overhang termini of only one or two nucleotides.

Highlights

• Recombinant enzyme
• Purified from non-animal host with a lower level of intrinsic RNases

Applications

• Preparation of DNA-free RNA
• Removal of template DNA following in vitro transcription
• Preparation of DNA-free RNA prior to RT-PCR and RT-qPCR
• DNA labeling by nick-translation in conjunction with DNA Polymerase I
• Studies of DNA-protein interactions by DNase I, RNase-free footprinting
• Generation of a library of randomly overlapping DNA inserts. Reaction buffer containing Mn2+ is used

Note

DNase I is sensitive to physical denaturation. Mix gently by inverting the tube. Do not vortex.

T4 Polynucleotide Kinase (10 U/µL) Thermo Scientific™

Thermo Scientific™ T4 Polynucleotide Kinase (T4 PNK) catalyzes the transfer of the gamma-phosphate from ATP to the 5'-OH group of single- and double-stranded DNAs and RNAs, oligonucleotides, or nucleoside 3'-monophosphates (forward reaction). The reaction is reversible. In the presence of ADP, T4 Polynucleotide Kinase exhibits 5'-phosphatase activity and catalyzes the exchange of phosphate groups between 5'-P-oligo-polynucleotides and ATP (exchange reaction). The enzyme is also a 3'-phosphatase.

Highlights
• Active in Thermo Scientific restriction enzyme, RT, and T4 DNA ligase buffers

Applications
• Labeling 5' -termini of nucleic acids to be used as:
    --Probes for hybridization
    --Probes for transcript mapping
    --Markers for gel electrophoresis
    --Primers for DNA sequencing
    --Primers for PCR
• 5'-phosphorylation of oligonucleotides, PCR products, other DNA or RNA prior to ligation
• Phosphorylation of PCR primers
• Detection of DNA modification by the [32P]-postlabeling assay
• Removal of 3'-phosphate groups

Notes
• The 5'-termini of nucleic acids can be labeled by either the forward or the exchange reaction.
• Polyethylene glycol (PEG) and spermidine improve the rate and efficiency of the phosphorylation reaction. PEG is used in the exchange reaction mixture.
• Since T4 Polynucleotide Kinase is inhibited by ammonium ions, use sodium acetate to precipitate DNA prior to phosphorylation.

Endonuclease V, T.maritima (5 U/µL) Thermo Scientific™

Thermo Scientific Endonuclease V, T. maritima (Endo V) is a 3'-endonuclease involved in DNA repair, which initiates removal of deaminated bases from damaged DNA, including uracil, hypoxanthine, and xanthine.

Endonuclease V is also active toward abasic sites and urea sites, base pair mismatches, flap and pseudo Y structures, and small insertions/deletions in DNA molecules. The cleavage site generated by Endonuclease V is at the second phosphodiester bond 3' to a lesion.

Highlights

• Optimal activity at temperatures of 65 to 70°C

Applications

• High-throughput methods for mutation research
• Studies in mutagenesis and DNA repair
• Mismatch cleavage
• Genotyping

Note

Use of this enzyme in certain applications may be covered by patents and may require a license.

When the enzyme is in excess, the primary nicked products experience a second nicking event on the complementary strand, leading to a double-stranded break. At low concentrations, however, Endonuclease V first nicks a DNA strand at the lesions located closer to the 5'-end of DNA molecule. Single-stranded DNA is cleaved with much lower efficiency. Mg2+ or Mn2+ ions are required for enzyme activity.

Topoisomerase I Invitrogen™

Topoisomerase I (DNA-relaxing enzyme) catalyzes the removal of superhelical turns from covalently closed DNA by a transient breakage and rejoining of phosphodiester bonds. Topoisomerase I is active in the presence of EDTA.

Applications: Relaxing positively and negatively supercoiled DNA (1). Producing DNA topoisomers (2).

Source: Purified from calf thymus.

Performance and Quality Testing: Endodeoxyribonuclease, 3´ and 5´ exodeoxyribonuclease, and phosphatase assays; conversion of super-coiled DNA to relaxed DNA.

Unit Definition: One unit catalyzes the conversion of 0.5 µg of superhelical Φ X174 RF DNA to a relaxed state in 30 min. at 37°C.

Unit Reaction Conditions: 50 mM Tris-HCl (pH 7.5), 50 mM KCl, 10 mM MgCl2 , 0.1 mM EDTA, 0.5 mM DTT, 30 µg/ml/BSA, 0.5 µg
Φ X174 RF DNA, and enzyme in 50 µl for 30 min. at 37°C.

ITK Recombinant Human Protein Thermo Scientific™

ITK is a tyrosine kinase expressed in T-cells which contains both SH2 and SH3 domains. ITK is thought to play a role in T-cell proliferation and differentiation.

CYP3A4 BACULOSOMES™ Plus Reagent, rHuman Invitrogen™

Cytochrome P450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells infected with recombinant baculovirus containing a human CYP450 isozyme, as well as human cytochrome P450 reductase. For this particular isozyme (CYP3A4), human cytochrome b5 is also included.
• Single human P450 isozyme for detailed drug metabolism studies
• High specific activity increases dynamic range and sensitivity
• No background metabolism for clear results

Single Overexpressed Human CYP450 Isozyme
CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over Human Liver Microsomes (HLMs) in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s or other classes of drug metabolizing enzymes.

High Activity Results in Increased Sensitivity
Compound rankings based on metabolism and inhibition profiles observed with CYP450 BACULOSOMES® Plus Reagents are very similar to those seen with HLMs for most compounds tested. However, CYP450 BACULOSOMES® Plus Reagents activity and metabolic rates with most substrates is significantly higher than those seen with HLMs. This results in a broad dynamic range and high sensitivity in assays utilizing CYP450 BACULOSOMES® Plus Reagents. The high level of activity and reproducibility of the enzyme component observed in the reaction with most probe substrates makes CYP450 BACULOSOMES® Plus Reagents well suited to high-throughput screening formats.

High Signal to Noise Ratio
No background metabolism by endogenous insect CYP450s has been detected with any of the Vivid® probe substrates tested so far.

Applications: isozyme identification, DMPK analysis, isozyme-specific metabolism, and inhibition screening

For Research Use Only. Not for any animal or human therapeutic or diagnostic use.
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