neb buffer 2  (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:
    NEBuffer 2 1
    Description:
    NEBuffer 2 1 5 0 ml
    Catalog Number:
    b7202s
    Price:
    24
    Size:
    5 0 ml
    Category:
    Buffers
    Buy from Supplier


    Structured Review

    New England Biolabs neb buffer 2
    NEBuffer 2 1
    NEBuffer 2 1 5 0 ml
    https://www.bioz.com/result/neb buffer 2/product/New England Biolabs
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    neb buffer 2 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference"

    Article Title: Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference

    Journal: Scientific Reports

    doi: 10.1038/srep09747

    Determination of divalent cation cofactor requirement and DNA substrate preference for GmrSD digestion. (A). Metal ion cofactor requirement for GmrSD digestion. Divalent cations or EDTA are indicated on top of each lane. (B). Substrate preference and optimal substrate size for GmrSD digestion. PCR DNA substrates containing 5hmC or regular dC were generated by PCR using pBR322 template and digested by GmrSD endonuclease in the presence of 1 mM ATP in NEB buffer 2. The same DNA substrates were also digested by HpaII (CCGG) in NEB buffer 4 (to confirm modified DNA).
    Figure Legend Snippet: Determination of divalent cation cofactor requirement and DNA substrate preference for GmrSD digestion. (A). Metal ion cofactor requirement for GmrSD digestion. Divalent cations or EDTA are indicated on top of each lane. (B). Substrate preference and optimal substrate size for GmrSD digestion. PCR DNA substrates containing 5hmC or regular dC were generated by PCR using pBR322 template and digested by GmrSD endonuclease in the presence of 1 mM ATP in NEB buffer 2. The same DNA substrates were also digested by HpaII (CCGG) in NEB buffer 4 (to confirm modified DNA).

    Techniques Used: Polymerase Chain Reaction, Generated, Modification

    Related Articles

    Gas Chromatography:

    Article Title: Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference
    Article Snippet: .. GmrSD enzyme activity assay T4 (glc-5hmC), T4gt (5hmC), and λ DNA (Dam+ Dcm+ ) or 5hmC-modified PCR DNA were digested with purified GmrSD enzyme in NEB buffer 2 (50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, 1 mM DTT) supplemented with 1 mM ATP at 37°C for 1 h unless specified otherwise. .. To generate 5hmC-modified PCR DNA substrates (266 bp, 0.5 kb, 1.0 kb, 1.9 kb, 3.8 kb), 5hm-dCTP (Zymo Research) was incorporated into PCR DNA by Taq DNA polymerase during PCR reactions.

    Agarose Gel Electrophoresis:

    Article Title: Rapid generation of incremental truncation libraries for protein engineering using ?-phosphothioate nucleotides
    Article Snippet: .. Twenty micrograms of pDIM-PGX in 200 µl of NEB buffer 2 (1× = 10 mM Tris–HCl pH 7.9, 10 mM MgCl2 , 100 mM NaCl, 1 mM DTT) were linearized by restriction digestion with Hin dIII (60 U) and purified by agarose gel electrophoresis. .. Three micrograms of linearized DNA were mixed with 6 µl of 10× exonuclease III buffer (Promega) and the volume adjusted to 60 µl with water.

    Article Title: DNA damage by C1027 involves hydrogen atom abstraction and addition to nucleobases
    Article Snippet: .. A 100 μL reaction containing pBR322 plasmid DNA (~10–15 μg), NEB buffer 2 (10 mM Tris-HCl, 10 mM MgCl2 , 50 mM NaCl, 1 mM DTT, pH 7.9), HindIII (3 μL, 20,000 U/mL, 60 units) and EcoRV (1 μL, 20,000 U/mL, 20 units) restriction enzymes were incubated at 37 °C for 1 h. Xylene cyanol dye was added to the reaction and ran down on a 1.8% agarose gel to separate fragments. .. The band corresponding to the 158 bp fragment was excised and purified using a QIAquick Gel Extraction Kit (Qiagen) and eluted with 30 μL of the provided elution buffer.

    Ligation:

    Article Title: ALS IMPLICATED PROTEIN TDP-43 SUSTAINS LEVELS OF STMN2 A MEDIATOR OF MOTOR NEURON GROWTH AND REPAIR
    Article Snippet: .. The annealed oligos were subsequently diluted 1:100 and 2 μL was added to the ligation reaction containing 2 μL of the 100 μM pUC6 vector, 2 μL of NEB buffer 2.1, 1 μL of 10mM DTT, 1 μL of 10mM ATP, 1 μL of BbsI (New England Biolabs), 0.5 μL of T7 ligase (New England Biolabs) and 10.5 μL of H2O. .. This solution was incubated in a thermocycler with the following cycle, 37°C for 5 minutes followed by 21°C for 5 minutes, repeated a total of 6 times.

    Enzyme Activity Assay:

    Article Title: Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference
    Article Snippet: .. GmrSD enzyme activity assay T4 (glc-5hmC), T4gt (5hmC), and λ DNA (Dam+ Dcm+ ) or 5hmC-modified PCR DNA were digested with purified GmrSD enzyme in NEB buffer 2 (50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, 1 mM DTT) supplemented with 1 mM ATP at 37°C for 1 h unless specified otherwise. .. To generate 5hmC-modified PCR DNA substrates (266 bp, 0.5 kb, 1.0 kb, 1.9 kb, 3.8 kb), 5hm-dCTP (Zymo Research) was incorporated into PCR DNA by Taq DNA polymerase during PCR reactions.

    Purification:

    Article Title: Rapid generation of incremental truncation libraries for protein engineering using ?-phosphothioate nucleotides
    Article Snippet: .. Twenty micrograms of pDIM-PGX in 200 µl of NEB buffer 2 (1× = 10 mM Tris–HCl pH 7.9, 10 mM MgCl2 , 100 mM NaCl, 1 mM DTT) were linearized by restriction digestion with Hin dIII (60 U) and purified by agarose gel electrophoresis. .. Three micrograms of linearized DNA were mixed with 6 µl of 10× exonuclease III buffer (Promega) and the volume adjusted to 60 µl with water.

    Article Title: Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference
    Article Snippet: .. GmrSD enzyme activity assay T4 (glc-5hmC), T4gt (5hmC), and λ DNA (Dam+ Dcm+ ) or 5hmC-modified PCR DNA were digested with purified GmrSD enzyme in NEB buffer 2 (50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, 1 mM DTT) supplemented with 1 mM ATP at 37°C for 1 h unless specified otherwise. .. To generate 5hmC-modified PCR DNA substrates (266 bp, 0.5 kb, 1.0 kb, 1.9 kb, 3.8 kb), 5hm-dCTP (Zymo Research) was incorporated into PCR DNA by Taq DNA polymerase during PCR reactions.

    Incubation:

    Article Title: DNA damage by C1027 involves hydrogen atom abstraction and addition to nucleobases
    Article Snippet: .. A 100 μL reaction containing pBR322 plasmid DNA (~10–15 μg), NEB buffer 2 (10 mM Tris-HCl, 10 mM MgCl2 , 50 mM NaCl, 1 mM DTT, pH 7.9), HindIII (3 μL, 20,000 U/mL, 60 units) and EcoRV (1 μL, 20,000 U/mL, 20 units) restriction enzymes were incubated at 37 °C for 1 h. Xylene cyanol dye was added to the reaction and ran down on a 1.8% agarose gel to separate fragments. .. The band corresponding to the 158 bp fragment was excised and purified using a QIAquick Gel Extraction Kit (Qiagen) and eluted with 30 μL of the provided elution buffer.

    Activity Assay:

    Article Title: Crystal structure of the modification-dependent SRA-HNH endonuclease TagI
    Article Snippet: .. Plasmid based assays TagI restriction activity was first assessed by digestion of 0.5–1 μg of pBR322 (Dcm+ or Dcm− ), pBRFM+ (Dcm+ ) or pACYC-HpyCH4IVM+ plasmid in NEB buffer 2.1 (50 mM NaCl, 10 mM Tris–HCl, 10 mM MgCl2 , 100 μg/ml BSA, pH 7.9) at 37°C for 1 h. After the reaction Protease K (1.6 U) was added at 37°C for 15 min. ..

    Article Title: Crystal structure of the modification-dependent SRA-HNH endonuclease TagI
    Article Snippet: .. TagI restriction activity was first assessed by digestion of 0.5–1 μg of pBR322 (Dcm+ or Dcm− ), pBRFM+ (Dcm+ ) or pACYC-HpyCH4IVM+ plasmid in NEB buffer 2.1 (50 mM NaCl, 10 mM Tris–HCl, 10 mM MgCl2 , 100 μg/ml BSA, pH 7.9) at 37°C for 1 h. After the reaction Protease K (1.6 U) was added at 37°C for 15 min. ..

    Polymerase Chain Reaction:

    Article Title: Expression and purification of a single-chain Type IV restriction enzyme Eco94GmrSD and determination of its substrate preference
    Article Snippet: .. GmrSD enzyme activity assay T4 (glc-5hmC), T4gt (5hmC), and λ DNA (Dam+ Dcm+ ) or 5hmC-modified PCR DNA were digested with purified GmrSD enzyme in NEB buffer 2 (50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, 1 mM DTT) supplemented with 1 mM ATP at 37°C for 1 h unless specified otherwise. .. To generate 5hmC-modified PCR DNA substrates (266 bp, 0.5 kb, 1.0 kb, 1.9 kb, 3.8 kb), 5hm-dCTP (Zymo Research) was incorporated into PCR DNA by Taq DNA polymerase during PCR reactions.

    Plasmid Preparation:

    Article Title: ALS IMPLICATED PROTEIN TDP-43 SUSTAINS LEVELS OF STMN2 A MEDIATOR OF MOTOR NEURON GROWTH AND REPAIR
    Article Snippet: .. The annealed oligos were subsequently diluted 1:100 and 2 μL was added to the ligation reaction containing 2 μL of the 100 μM pUC6 vector, 2 μL of NEB buffer 2.1, 1 μL of 10mM DTT, 1 μL of 10mM ATP, 1 μL of BbsI (New England Biolabs), 0.5 μL of T7 ligase (New England Biolabs) and 10.5 μL of H2O. .. This solution was incubated in a thermocycler with the following cycle, 37°C for 5 minutes followed by 21°C for 5 minutes, repeated a total of 6 times.

    Article Title: Crystal structure of the modification-dependent SRA-HNH endonuclease TagI
    Article Snippet: .. Plasmid based assays TagI restriction activity was first assessed by digestion of 0.5–1 μg of pBR322 (Dcm+ or Dcm− ), pBRFM+ (Dcm+ ) or pACYC-HpyCH4IVM+ plasmid in NEB buffer 2.1 (50 mM NaCl, 10 mM Tris–HCl, 10 mM MgCl2 , 100 μg/ml BSA, pH 7.9) at 37°C for 1 h. After the reaction Protease K (1.6 U) was added at 37°C for 15 min. ..

    Article Title: Crystal structure of the modification-dependent SRA-HNH endonuclease TagI
    Article Snippet: .. TagI restriction activity was first assessed by digestion of 0.5–1 μg of pBR322 (Dcm+ or Dcm− ), pBRFM+ (Dcm+ ) or pACYC-HpyCH4IVM+ plasmid in NEB buffer 2.1 (50 mM NaCl, 10 mM Tris–HCl, 10 mM MgCl2 , 100 μg/ml BSA, pH 7.9) at 37°C for 1 h. After the reaction Protease K (1.6 U) was added at 37°C for 15 min. ..

    Article Title: DNA damage by C1027 involves hydrogen atom abstraction and addition to nucleobases
    Article Snippet: .. A 100 μL reaction containing pBR322 plasmid DNA (~10–15 μg), NEB buffer 2 (10 mM Tris-HCl, 10 mM MgCl2 , 50 mM NaCl, 1 mM DTT, pH 7.9), HindIII (3 μL, 20,000 U/mL, 60 units) and EcoRV (1 μL, 20,000 U/mL, 20 units) restriction enzymes were incubated at 37 °C for 1 h. Xylene cyanol dye was added to the reaction and ran down on a 1.8% agarose gel to separate fragments. .. The band corresponding to the 158 bp fragment was excised and purified using a QIAquick Gel Extraction Kit (Qiagen) and eluted with 30 μL of the provided elution buffer.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs bst dna polymerase
    Single factor experiment of RealAmp. The influence of each variable on the LAMP reaction was analyzed by the amplification curve ( a ) and the melt peak ( b ). For the <t>Bst</t> <t>DNA</t> polymerase optimization, Bst polymerase quantities were adjusted to 2 U, 4 U, 6
    Bst Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bst dna polymerase/product/New England Biolabs
    Average 99 stars, based on 93 article reviews
    Price from $9.99 to $1999.99
    bst dna polymerase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    New England Biolabs t4 rnl2 reaction buffer
    Scheme of the splinted-ligation method in bRNA construction. In this method, a 2΄-5΄ linked ribo-guanosine (G)-nucleoside in an RNA strand containing the 5΄-segment and 2΄-arm (precursor 1) is transformed into a branchpoint nucleotide by ligation to an RNA strand representing the 3΄-arm (precursor 2). To do so, the two precursors are hybridized partially to a complementary RNA bridge. In this way, the 5΄-phosphate of precursor 2 is brought close to the free 3΄-hydroxyl of the 2΄-5΄ linked nucleoside of precursor 1. The two oligonucleotides are then joined by T4 RNA Ligase 2. Red, blue, and pink symbols ‘w’ represent RNA; the black line represents DNA. The 2΄-5΄ linked ribo-G-nucleoside in precursor 1 at nucleotide (nt) position 37 is highlighted. Nucleic acids downstream of a 2΄-5΄ linkage are plotted vertically in linear and branched oligonucleotides.
    T4 Rnl2 Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 rnl2 reaction buffer/product/New England Biolabs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t4 rnl2 reaction buffer - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    99
    New England Biolabs haeiii
    The mutational effects in the laboratory drift correlate with sequence exchanges in <t>M.HaeIII</t> orthologs. a . The positional rates of evolution in M.HaeIII’s natural orthologs (‘Rate4site’, μ; red line) were plotted alongside the positional W rel values in M.HaeIII (blue line). The positional W rel values correspond to the average W rel ∑ i { W r e l i ⋅ log 2 [ 1 + 10 ⋅ f ( G 17 i ) ] } , Where i refers to all the possible single nucleotide mutations at a given residue position. Upper panel –positions 2 to 175; Lower panel –positions 176 to 330. Noted are M.HaeIII’s key functional residues, those of the cofactor binding site, the catalytic residues including the enzyme’s reaction center (Cys71, black arrow), and the target <t>DNA</t> binding residues. Also noted are positions of compensatory mutations that were enriched in the drift W rel > 1.1, listed in S2 Table ). b. M.HaeIII’s three-dimensional structure illustrated as a cartoon (PDB id 1dct). Residues are colored from blue to red according to their averaged W rel values (as in c ). The cofactor, SAM, is in yellow, and the enzyme’s catalytic residue (Cys71) is in green. c. The same as b for the positional diversity calculated by Rate4site (μ, as in c ) [ 65 ]. d. The distribution of PROVEAN scores for all possible single nucleotide missense mutations (n = 1,957). Shown are the distribution of mutations categorized as ‘deleterious’ ( W rel ≤0.6), and of mutations categorized as ‘nearly-neutral’, ‘neutral’ and ‘beneficial’ ( W rel > 0.6). e. The same distribution while excluding ‘nearly-neutral’ mutations.
    Haeiii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 84 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/haeiii/product/New England Biolabs
    Average 99 stars, based on 84 article reviews
    Price from $9.99 to $1999.99
    haeiii - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Single factor experiment of RealAmp. The influence of each variable on the LAMP reaction was analyzed by the amplification curve ( a ) and the melt peak ( b ). For the Bst DNA polymerase optimization, Bst polymerase quantities were adjusted to 2 U, 4 U, 6

    Journal: Scientific Reports

    Article Title: Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Ustilago maydis

    doi: 10.1038/s41598-017-13881-4

    Figure Lengend Snippet: Single factor experiment of RealAmp. The influence of each variable on the LAMP reaction was analyzed by the amplification curve ( a ) and the melt peak ( b ). For the Bst DNA polymerase optimization, Bst polymerase quantities were adjusted to 2 U, 4 U, 6

    Article Snippet: This optimized reaction mixture (total, 25 μl) contained 12.5 μl 2 × Bst DNA polymerase (NEB, Ipswich, MA, USA) reaction buffer [1 × containing 1.6 mM dNTPs, 1 M betaine, 4 mM MgSO4 , 20 mM Tris-HCl (pH 8.8), 10 mM KCl, 10 mM (NH4)2 SO4 , and 0.1% Triton X-20 (Sigma-Aldrich Inc., Saint Louis, USA), Double Helix Tech.

    Techniques: Amplification

    Scheme of the splinted-ligation method in bRNA construction. In this method, a 2΄-5΄ linked ribo-guanosine (G)-nucleoside in an RNA strand containing the 5΄-segment and 2΄-arm (precursor 1) is transformed into a branchpoint nucleotide by ligation to an RNA strand representing the 3΄-arm (precursor 2). To do so, the two precursors are hybridized partially to a complementary RNA bridge. In this way, the 5΄-phosphate of precursor 2 is brought close to the free 3΄-hydroxyl of the 2΄-5΄ linked nucleoside of precursor 1. The two oligonucleotides are then joined by T4 RNA Ligase 2. Red, blue, and pink symbols ‘w’ represent RNA; the black line represents DNA. The 2΄-5΄ linked ribo-G-nucleoside in precursor 1 at nucleotide (nt) position 37 is highlighted. Nucleic acids downstream of a 2΄-5΄ linkage are plotted vertically in linear and branched oligonucleotides.

    Journal: Nucleic Acids Research

    Article Title: Arm-specific cleavage and mutation during reverse transcription of 2΄,5΄-branched RNA by Moloney murine leukemia virus reverse transcriptase

    doi: 10.1093/nar/gkx073

    Figure Lengend Snippet: Scheme of the splinted-ligation method in bRNA construction. In this method, a 2΄-5΄ linked ribo-guanosine (G)-nucleoside in an RNA strand containing the 5΄-segment and 2΄-arm (precursor 1) is transformed into a branchpoint nucleotide by ligation to an RNA strand representing the 3΄-arm (precursor 2). To do so, the two precursors are hybridized partially to a complementary RNA bridge. In this way, the 5΄-phosphate of precursor 2 is brought close to the free 3΄-hydroxyl of the 2΄-5΄ linked nucleoside of precursor 1. The two oligonucleotides are then joined by T4 RNA Ligase 2. Red, blue, and pink symbols ‘w’ represent RNA; the black line represents DNA. The 2΄-5΄ linked ribo-G-nucleoside in precursor 1 at nucleotide (nt) position 37 is highlighted. Nucleic acids downstream of a 2΄-5΄ linkage are plotted vertically in linear and branched oligonucleotides.

    Article Snippet: The ligation reaction was performed in 20 μl containing 1× T4 Rnl2 reaction buffer (NEB), 12.5% (w/v) polyethylene glycol (PEG) 8000 or PEG 4000 and 5 units of T4 RNA Ligase 2 (NEB).

    Techniques: Ligation, Transformation Assay

    The mutational effects in the laboratory drift correlate with sequence exchanges in M.HaeIII orthologs. a . The positional rates of evolution in M.HaeIII’s natural orthologs (‘Rate4site’, μ; red line) were plotted alongside the positional W rel values in M.HaeIII (blue line). The positional W rel values correspond to the average W rel ∑ i { W r e l i ⋅ log 2 [ 1 + 10 ⋅ f ( G 17 i ) ] } , Where i refers to all the possible single nucleotide mutations at a given residue position. Upper panel –positions 2 to 175; Lower panel –positions 176 to 330. Noted are M.HaeIII’s key functional residues, those of the cofactor binding site, the catalytic residues including the enzyme’s reaction center (Cys71, black arrow), and the target DNA binding residues. Also noted are positions of compensatory mutations that were enriched in the drift W rel > 1.1, listed in S2 Table ). b. M.HaeIII’s three-dimensional structure illustrated as a cartoon (PDB id 1dct). Residues are colored from blue to red according to their averaged W rel values (as in c ). The cofactor, SAM, is in yellow, and the enzyme’s catalytic residue (Cys71) is in green. c. The same as b for the positional diversity calculated by Rate4site (μ, as in c ) [ 65 ]. d. The distribution of PROVEAN scores for all possible single nucleotide missense mutations (n = 1,957). Shown are the distribution of mutations categorized as ‘deleterious’ ( W rel ≤0.6), and of mutations categorized as ‘nearly-neutral’, ‘neutral’ and ‘beneficial’ ( W rel > 0.6). e. The same distribution while excluding ‘nearly-neutral’ mutations.

    Journal: PLoS Computational Biology

    Article Title: Systematic Mapping of Protein Mutational Space by Prolonged Drift Reveals the Deleterious Effects of Seemingly Neutral Mutations

    doi: 10.1371/journal.pcbi.1004421

    Figure Lengend Snippet: The mutational effects in the laboratory drift correlate with sequence exchanges in M.HaeIII orthologs. a . The positional rates of evolution in M.HaeIII’s natural orthologs (‘Rate4site’, μ; red line) were plotted alongside the positional W rel values in M.HaeIII (blue line). The positional W rel values correspond to the average W rel ∑ i { W r e l i ⋅ log 2 [ 1 + 10 ⋅ f ( G 17 i ) ] } , Where i refers to all the possible single nucleotide mutations at a given residue position. Upper panel –positions 2 to 175; Lower panel –positions 176 to 330. Noted are M.HaeIII’s key functional residues, those of the cofactor binding site, the catalytic residues including the enzyme’s reaction center (Cys71, black arrow), and the target DNA binding residues. Also noted are positions of compensatory mutations that were enriched in the drift W rel > 1.1, listed in S2 Table ). b. M.HaeIII’s three-dimensional structure illustrated as a cartoon (PDB id 1dct). Residues are colored from blue to red according to their averaged W rel values (as in c ). The cofactor, SAM, is in yellow, and the enzyme’s catalytic residue (Cys71) is in green. c. The same as b for the positional diversity calculated by Rate4site (μ, as in c ) [ 65 ]. d. The distribution of PROVEAN scores for all possible single nucleotide missense mutations (n = 1,957). Shown are the distribution of mutations categorized as ‘deleterious’ ( W rel ≤0.6), and of mutations categorized as ‘nearly-neutral’, ‘neutral’ and ‘beneficial’ ( W rel > 0.6). e. The same distribution while excluding ‘nearly-neutral’ mutations.

    Article Snippet: About 106 individual transformants were obtained in each round. (ii ) Colonies grown at 37°C overnight were combined, plasmid DNA was extracted and digested with HaeIII (10–20 units, in 50 μl of NEB buffer 2, for 2 hours at 37°C), and re-purified (PCR purification kit, QIAGEN). (iii ) The recovered plasmid DNA was re-transformed for another round of enrichment.

    Techniques: Sequencing, Functional Assay, Binding Assay