user enzyme mix  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    USER Enzyme
    Description:
    USER Enzyme 250 units
    Catalog Number:
    m5505l
    Price:
    297
    Size:
    250 units
    Category:
    Other Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs user enzyme mix
    USER Enzyme
    USER Enzyme 250 units
    https://www.bioz.com/result/user enzyme mix/product/New England Biolabs
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    user enzyme mix - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Simultaneous and stoichiometric purification of hundreds of oligonucleotides"

    Article Title: Simultaneous and stoichiometric purification of hundreds of oligonucleotides

    Journal: Nature Communications

    doi: 10.1038/s41467-018-04870-w

    Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by streptavidin-coated magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the USER enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized
    Figure Legend Snippet: Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by streptavidin-coated magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the USER enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized

    Techniques Used: Purification, Synthesized, Sequencing, Concentration Assay, Oligo Synthesis, Magnetic Beads, Next-Generation Sequencing, Standard Deviation, Binding Assay

    Related Articles

    Clone Assay:

    Article Title: Small Molecule-Triggered Cas9 Protein with Improved Genome-Editing Specificity
    Article Snippet: .. Intein 37R3-2 was subcloned at the described positions into the wild-type Cas9 expression plasmid using USER (NEB M5505) cloning. sgRNA expression plasmids used in this study have been described previously . ..

    Amplification:

    Article Title: Inhibitors of MyD88-Dependent Proinflammatory Cytokine Production Identified Utilizing a Novel RNA Interference Screening Approach
    Article Snippet: .. Step 7 To generate cohesive ends, the amplified PCR product was treated with the USER enzyme (New England Biolabs) according to the manufacturer's instructions, which specifically removes deoxyuridine in the DNA. .. The final product contains unique 7 bp and 4 bp overhangs at each end, which were used to facilitate cloning into the lentiviral vector pLL3.7 (kindly provided by Dr. Luk Van Parijs).

    Article Title: Artifactual mutations resulting from DNA lesions limit detection levels in ultrasensitive sequencing applications
    Article Snippet: .. Treatments with the USER enzyme were performed on HSI_insert_1 by incubating 2 × 107 copies HSI_insert construct with 1 U USER enzyme (NEB) in 1× Phusion HF Buffer in a reaction volume of 20 µl at 37 °C for 30 min prior to amplification. .. For duplex sequencing, 10 µl purified, adapter ligated library of insert 3 were incubated with 1 U USER enzyme in 1× NEB CS Buffer in a reaction volume of 20 µl at 37 °C for 1h.

    Magnetic Beads:

    Article Title: AutoRELACS: Automated Generation And Analysis Of Ultra-parallel ChIP-seq
    Article Snippet: .. The following reagents are required for this section of program, as specified in the instrument setup ( ): 100% isopropanol, EB (10 mM Tris-HCl pH 8), freshly prepared 85% ethanol (all on the deck at room temperature), proteinase K 20 mg/ml (Thermo Fisher, EO0491), glycogen 20 mg/mg (Thermo Fisher, R0561), carboxylated magnetic beads (Invitrogen, 65011), PCR mix (NEBNext Ultra II Q5 Master mix, NEB M0544), USER enzyme (NEB M5505), all placed in 1.5 ml conical tubes in a cold Peltier block. .. Ampure XP (Beckman Coulter, A63881) are thoroughly mixed and aliquoted column-wise according to the pattern of “Sample Plate 2” in a 96-well storage plate (AB0765, Thermo Fisher), using 100 µl of beads per well.

    Construct:

    Article Title: Artifactual mutations resulting from DNA lesions limit detection levels in ultrasensitive sequencing applications
    Article Snippet: .. Treatments with the USER enzyme were performed on HSI_insert_1 by incubating 2 × 107 copies HSI_insert construct with 1 U USER enzyme (NEB) in 1× Phusion HF Buffer in a reaction volume of 20 µl at 37 °C for 30 min prior to amplification. .. For duplex sequencing, 10 µl purified, adapter ligated library of insert 3 were incubated with 1 U USER enzyme in 1× NEB CS Buffer in a reaction volume of 20 µl at 37 °C for 1h.

    Purification:

    Article Title: A versatile element for gene addition in bacterial chromosomes
    Article Snippet: .. General materials: ∘ USERBstBI-compatible digested pMS26 (from step 1) ∘ PfuCx_TurboCx _Hotstart_DNA_polymerase (Agilent Genomics) ∘ USER enzyme (NEB M5505) ∘ PCR purification columns ∘ Universal flanking primers (glmS, ptsS ) to monitor chromosomal insertion ∘ RB ampicillin plates ∘ RB no drug plates ∘ Incubators at 30°C and 42°C ∘ SOC or other outgrowth medium Experiment-specific materials: ∘ competent host cells ∘ DNA template ∘ gene-specific primers with 5′ sequences suitable to generate USERBstBI-compatible extensions. ..

    Incubation:

    Article Title: Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage
    Article Snippet: .. Following protein expression, 5 μL of lysate was combined with 35 μL of ssDNA (1.8 μM) and USER enzyme (1 unit) in CutSmart buffer (New England Biolabs) (50 mM potassium acetate, 29 mM Tris-acetate, 10 mM magnesium acetate, 100 ug/mL BSA, pH 7.9) and incubated at 37 °C for 2 h. Cleaved U-containing substrates were resolved from full-length unmodified substrates on a 10% TBE-urea gel (Bio-Rad). .. Expression and purification of His6 -rAPOBEC1-linker-dCas9 fusions E. coli BL21 STAR (DE3)-competent cells (ThermoFisher Scientific) were transformed with plasmids encoding pET28b-His6 -rAPOBEC-linker-dCas9 with GGS, (GGS)3 , XTEN, or (GGS)7 linkers.

    Blocking Assay:

    Article Title: AutoRELACS: Automated Generation And Analysis Of Ultra-parallel ChIP-seq
    Article Snippet: .. The following reagents are required for this section of program, as specified in the instrument setup ( ): 100% isopropanol, EB (10 mM Tris-HCl pH 8), freshly prepared 85% ethanol (all on the deck at room temperature), proteinase K 20 mg/ml (Thermo Fisher, EO0491), glycogen 20 mg/mg (Thermo Fisher, R0561), carboxylated magnetic beads (Invitrogen, 65011), PCR mix (NEBNext Ultra II Q5 Master mix, NEB M0544), USER enzyme (NEB M5505), all placed in 1.5 ml conical tubes in a cold Peltier block. .. Ampure XP (Beckman Coulter, A63881) are thoroughly mixed and aliquoted column-wise according to the pattern of “Sample Plate 2” in a 96-well storage plate (AB0765, Thermo Fisher), using 100 µl of beads per well.

    Expressing:

    Article Title: Small Molecule-Triggered Cas9 Protein with Improved Genome-Editing Specificity
    Article Snippet: .. Intein 37R3-2 was subcloned at the described positions into the wild-type Cas9 expression plasmid using USER (NEB M5505) cloning. sgRNA expression plasmids used in this study have been described previously . ..

    Article Title: Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage
    Article Snippet: .. Following protein expression, 5 μL of lysate was combined with 35 μL of ssDNA (1.8 μM) and USER enzyme (1 unit) in CutSmart buffer (New England Biolabs) (50 mM potassium acetate, 29 mM Tris-acetate, 10 mM magnesium acetate, 100 ug/mL BSA, pH 7.9) and incubated at 37 °C for 2 h. Cleaved U-containing substrates were resolved from full-length unmodified substrates on a 10% TBE-urea gel (Bio-Rad). .. Expression and purification of His6 -rAPOBEC1-linker-dCas9 fusions E. coli BL21 STAR (DE3)-competent cells (ThermoFisher Scientific) were transformed with plasmids encoding pET28b-His6 -rAPOBEC-linker-dCas9 with GGS, (GGS)3 , XTEN, or (GGS)7 linkers.

    Polymerase Chain Reaction:

    Article Title: Inhibitors of MyD88-Dependent Proinflammatory Cytokine Production Identified Utilizing a Novel RNA Interference Screening Approach
    Article Snippet: .. Step 7 To generate cohesive ends, the amplified PCR product was treated with the USER enzyme (New England Biolabs) according to the manufacturer's instructions, which specifically removes deoxyuridine in the DNA. .. The final product contains unique 7 bp and 4 bp overhangs at each end, which were used to facilitate cloning into the lentiviral vector pLL3.7 (kindly provided by Dr. Luk Van Parijs).

    Article Title: AutoRELACS: Automated Generation And Analysis Of Ultra-parallel ChIP-seq
    Article Snippet: .. The following reagents are required for this section of program, as specified in the instrument setup ( ): 100% isopropanol, EB (10 mM Tris-HCl pH 8), freshly prepared 85% ethanol (all on the deck at room temperature), proteinase K 20 mg/ml (Thermo Fisher, EO0491), glycogen 20 mg/mg (Thermo Fisher, R0561), carboxylated magnetic beads (Invitrogen, 65011), PCR mix (NEBNext Ultra II Q5 Master mix, NEB M0544), USER enzyme (NEB M5505), all placed in 1.5 ml conical tubes in a cold Peltier block. .. Ampure XP (Beckman Coulter, A63881) are thoroughly mixed and aliquoted column-wise according to the pattern of “Sample Plate 2” in a 96-well storage plate (AB0765, Thermo Fisher), using 100 µl of beads per well.

    Article Title: A versatile element for gene addition in bacterial chromosomes
    Article Snippet: .. General materials: ∘ USERBstBI-compatible digested pMS26 (from step 1) ∘ PfuCx_TurboCx _Hotstart_DNA_polymerase (Agilent Genomics) ∘ USER enzyme (NEB M5505) ∘ PCR purification columns ∘ Universal flanking primers (glmS, ptsS ) to monitor chromosomal insertion ∘ RB ampicillin plates ∘ RB no drug plates ∘ Incubators at 30°C and 42°C ∘ SOC or other outgrowth medium Experiment-specific materials: ∘ competent host cells ∘ DNA template ∘ gene-specific primers with 5′ sequences suitable to generate USERBstBI-compatible extensions. ..

    Plasmid Preparation:

    Article Title: Small Molecule-Triggered Cas9 Protein with Improved Genome-Editing Specificity
    Article Snippet: .. Intein 37R3-2 was subcloned at the described positions into the wild-type Cas9 expression plasmid using USER (NEB M5505) cloning. sgRNA expression plasmids used in this study have been described previously . ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs dpnii
    Scalable gene synthesis platform schematic for OLS Pool 2 Pre-designed oligonucleotides (no distinction is made between dsDNA and ssDNA in the figure) are synthesized on a DNA microchip ( a ) and then cleaved to make a pool of oligonucleotides (b) . Plate-specific primer sequences (yellow or brown) are used to amplify separate Plate Subpools (c) (only two are shown), which contain DNA to assemble different genes (only three are shown for each plate subpool). Assembly specific sequences (shades of blue) are used to amplify assembly subpools (d) that contain only the DNA required to make a single gene. The primer sequences are cleaved (e) using either Type IIS restriction enzymes (resulting in dsDNA) or by <t>DpnII/USER/γ</t> exonuclease processing (producing ssDNA). Construction primers (shown as white and black sites flanking the full assembly) are then used in an assembly PCR reaction to build a gene from each assembly subpool (f) . Depending on the downstream application the assembled products are then cloned either before or after an enzymatic error correction step.
    Dpnii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dpnii/product/New England Biolabs
    Average 99 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    dpnii - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    88
    New England Biolabs user enzyme mix
    Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by <t>streptavidin-coated</t> magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the <t>USER</t> enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized
    User Enzyme Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/user enzyme mix/product/New England Biolabs
    Average 88 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    user enzyme mix - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    99
    New England Biolabs dna glycosylase lyase endo viii
    Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by <t>streptavidin-coated</t> magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the <t>USER</t> enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized
    Dna Glycosylase Lyase Endo Viii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 951 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna glycosylase lyase endo viii/product/New England Biolabs
    Average 99 stars, based on 951 article reviews
    Price from $9.99 to $1999.99
    dna glycosylase lyase endo viii - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Scalable gene synthesis platform schematic for OLS Pool 2 Pre-designed oligonucleotides (no distinction is made between dsDNA and ssDNA in the figure) are synthesized on a DNA microchip ( a ) and then cleaved to make a pool of oligonucleotides (b) . Plate-specific primer sequences (yellow or brown) are used to amplify separate Plate Subpools (c) (only two are shown), which contain DNA to assemble different genes (only three are shown for each plate subpool). Assembly specific sequences (shades of blue) are used to amplify assembly subpools (d) that contain only the DNA required to make a single gene. The primer sequences are cleaved (e) using either Type IIS restriction enzymes (resulting in dsDNA) or by DpnII/USER/γ exonuclease processing (producing ssDNA). Construction primers (shown as white and black sites flanking the full assembly) are then used in an assembly PCR reaction to build a gene from each assembly subpool (f) . Depending on the downstream application the assembled products are then cloned either before or after an enzymatic error correction step.

    Journal: Nature biotechnology

    Article Title: A Scalable Gene Synthesis Platform Using High-Fidelity DNA Microchips

    doi: 10.1038/nbt.1716

    Figure Lengend Snippet: Scalable gene synthesis platform schematic for OLS Pool 2 Pre-designed oligonucleotides (no distinction is made between dsDNA and ssDNA in the figure) are synthesized on a DNA microchip ( a ) and then cleaved to make a pool of oligonucleotides (b) . Plate-specific primer sequences (yellow or brown) are used to amplify separate Plate Subpools (c) (only two are shown), which contain DNA to assemble different genes (only three are shown for each plate subpool). Assembly specific sequences (shades of blue) are used to amplify assembly subpools (d) that contain only the DNA required to make a single gene. The primer sequences are cleaved (e) using either Type IIS restriction enzymes (resulting in dsDNA) or by DpnII/USER/γ exonuclease processing (producing ssDNA). Construction primers (shown as white and black sites flanking the full assembly) are then used in an assembly PCR reaction to build a gene from each assembly subpool (f) . Depending on the downstream application the assembled products are then cloned either before or after an enzymatic error correction step.

    Article Snippet: PCR amplification was followed by γ exonuclease digestion of 5′ phosphorylated strands, hybridization of the 3′ primer site to its complement, and cleavage of the 5′ and 3′ primer sites using USER enzyme mix and DpnII (New England Biolabs), respectively.

    Techniques: Synthesized, MicroChIP Assay, Polymerase Cycling Assembly, Clone Assay

    Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by streptavidin-coated magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the USER enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized

    Journal: Nature Communications

    Article Title: Simultaneous and stoichiometric purification of hundreds of oligonucleotides

    doi: 10.1038/s41467-018-04870-w

    Figure Lengend Snippet: Stoichiometrically normalizing oligonucleotide purification (SNOP) concept and workflow. a The input reagents for SNOP are chemically synthesized oligonucleotide precursors P 1 through P N that contain imperfect synthesis products with 5′ truncations and/or internal deletions, and with potentially very different concentrations. SNOP produces a pool of oligonucleotide products O 1 through O N that has high fractions of oligos with perfect sequence, and with all products at roughly equal concentration. SNOP uses a single biotinylated capture probe oligonucleotide synthesized with a degenerate “SWSWSW” randomer subsequence. Each instance of the randomer is complementary to one precursor tag sequence. The different instances of the capture probe are all at roughly equal concentration, due to split-pool oligo synthesis. Precursors with perfect tag sequences hybridize to the probe and are captured by streptavidin-coated magnetic beads. Subsequent cleavage at the deoxyuracil (dU) site using the USER enzyme mix ( https://www.neb.com/products/m5505-user-enzyme ) releases the oligo products into solution. Setting the capture probe to be the limiting reagent allows all SNOP products to be all at roughly equal concentrations. b SNOP enriches the fraction of perfect oligos because synthesis errors are correlated; molecules with no truncations or deletions in the tag sequences are also more likely to not have any deletions in the oligo product sequence. Shown in this panel are NGS sequence analysis results of a pool of N = 64 precursor oligonucleotides; error bars show standard deviation across different oligos (see Methods for library preparation details). c SNOP is very sensitive to small sequence changes in the tag; even single-nucleotide variations result in significantly reduced binding yield (see also Supplementary Note). This property allows SNOP products to be both highly pure and stoichiometrically normalized

    Article Snippet: Subsequent solid-phase separation using streptavidin-coated magnetic beads removes unbound precursors, and applying USER enzyme mix (New England Biolabs) cleaves the oligo products from the tags at the dU site.

    Techniques: Purification, Synthesized, Sequencing, Concentration Assay, Oligo Synthesis, Magnetic Beads, Next-Generation Sequencing, Standard Deviation, Binding Assay