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    Name:
    BbvCI
    Description:
    BbvCI 500 units
    Catalog Number:
    r0601l
    Price:
    269
    Size:
    500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs bbvci
    BbvCI
    BbvCI 500 units
    https://www.bioz.com/result/bbvci/product/New England Biolabs
    Average 94 stars, based on 43 article reviews
    Price from $9.99 to $1999.99
    bbvci - by Bioz Stars, 2020-07
    94/100 stars

    Images

    1) Product Images from "Structural diversity of supercoiled DNA"

    Article Title: Structural diversity of supercoiled DNA

    Journal: Nature Communications

    doi: 10.1038/ncomms9440

    Effect of supercoiling on the structure of minicircle DNA. ( a ) Individual 336 bp minicircle topoisomers were isolated and analysed by polyacrylamide gel electrophoresis in the presence of 10 mM CaCl 2 . Mr: 100 bp DNA ladder, L: minicircle linearized by EcoRV, N: minicircle nicked by Nb.BbvCI. ( b ) Projections of cryo-ET subtomograms of hydrated 336 bp DNA minicircles of the Lk =34 topoisomer. ( c ) Commonly observed shapes were open circle, open figure-8, figure-8, racquet, handcuffs, needle, and rod, each of which are shown in orthogonal views. ( d ) Other shapes observed, especially in the more highly supercoiled topoisomers. ( e ) Shape frequency distribution plot for each topoisomer population (n=number of minicircles analysed). A weighted average for each topoisomer, approximating the average degree of compactness, is denoted by the black triangle. The weighted average was calculated by assigning each conformation a value that increased in line with compactness. Open circles were given a value of 1, open figure-8 s a value of 2, figure-8 s as a value of 3, and so on. The relative fraction of each was subsequently used to determine the average degree of compactness. Lk , Δ Lk and superhelical density (σ) for each topoisomer are shown (see Supplementary Note 1 ).
    Figure Legend Snippet: Effect of supercoiling on the structure of minicircle DNA. ( a ) Individual 336 bp minicircle topoisomers were isolated and analysed by polyacrylamide gel electrophoresis in the presence of 10 mM CaCl 2 . Mr: 100 bp DNA ladder, L: minicircle linearized by EcoRV, N: minicircle nicked by Nb.BbvCI. ( b ) Projections of cryo-ET subtomograms of hydrated 336 bp DNA minicircles of the Lk =34 topoisomer. ( c ) Commonly observed shapes were open circle, open figure-8, figure-8, racquet, handcuffs, needle, and rod, each of which are shown in orthogonal views. ( d ) Other shapes observed, especially in the more highly supercoiled topoisomers. ( e ) Shape frequency distribution plot for each topoisomer population (n=number of minicircles analysed). A weighted average for each topoisomer, approximating the average degree of compactness, is denoted by the black triangle. The weighted average was calculated by assigning each conformation a value that increased in line with compactness. Open circles were given a value of 1, open figure-8 s a value of 2, figure-8 s as a value of 3, and so on. The relative fraction of each was subsequently used to determine the average degree of compactness. Lk , Δ Lk and superhelical density (σ) for each topoisomer are shown (see Supplementary Note 1 ).

    Techniques Used: Isolation, Polyacrylamide Gel Electrophoresis

    2) Product Images from "Cleavage of a model DNA replication fork by a Type I restriction endonuclease"

    Article Title: Cleavage of a model DNA replication fork by a Type I restriction endonuclease

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkp214

    Preparation of a long branched DNA (eF1). ( A ) pES1 was treated with SphI, PvuII and nicking endonuclease Nb.BbvCI. A partial fragment from pGap1 was prepared by PCR with indicated primers, followed by nicking with Nt.BbvCI. ( B ) Single strands between the nicks introduced in the previous step were dissociated by heating with removal by complementary oligo DNA from the fragment containing a gap structure. ( C ) Resulting DNAs with the gap were annealed to form a branched structure (Materials and methods section). A leftward triangle indicates an EcoR124I site, 5′ CGA TGCTGTA TTC . An open circle indicates 32 P for 5′-end labeling.
    Figure Legend Snippet: Preparation of a long branched DNA (eF1). ( A ) pES1 was treated with SphI, PvuII and nicking endonuclease Nb.BbvCI. A partial fragment from pGap1 was prepared by PCR with indicated primers, followed by nicking with Nt.BbvCI. ( B ) Single strands between the nicks introduced in the previous step were dissociated by heating with removal by complementary oligo DNA from the fragment containing a gap structure. ( C ) Resulting DNAs with the gap were annealed to form a branched structure (Materials and methods section). A leftward triangle indicates an EcoR124I site, 5′ CGA TGCTGTA TTC . An open circle indicates 32 P for 5′-end labeling.

    Techniques Used: Polymerase Chain Reaction, End Labeling

    3) Product Images from "A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens"

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens

    Journal: Plant Methods

    doi: 10.1186/1746-4811-7-42

    Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.
    Figure Legend Snippet: Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.

    Techniques Used: Clone Assay, Expressing, Plasmid Preparation, Selection

    The destination vector pEG101-SacB/R can be stably propagated in the E. coli strain DH5α . Restriction digestion ( Bst XI and Bbv CI) of plasmid DNAs, isolated from five repeated overnight cultures of E. coli carrying the pEG101-SacB/R destination vector, showed identical restriction patterns. M: 1 kb marker. Lanes 1 through 5 were digested plasmid DNAs of five repeated subcultures.
    Figure Legend Snippet: The destination vector pEG101-SacB/R can be stably propagated in the E. coli strain DH5α . Restriction digestion ( Bst XI and Bbv CI) of plasmid DNAs, isolated from five repeated overnight cultures of E. coli carrying the pEG101-SacB/R destination vector, showed identical restriction patterns. M: 1 kb marker. Lanes 1 through 5 were digested plasmid DNAs of five repeated subcultures.

    Techniques Used: Plasmid Preparation, Stable Transfection, Isolation, Marker

    4) Product Images from "Sensitive isothermal detection of nucleic-acid sequence by primer generation-rolling circle amplification"

    Article Title: Sensitive isothermal detection of nucleic-acid sequence by primer generation-rolling circle amplification

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkn1014

    Endpoint product analysis of PG–RCA. ( A ) Endpoint products in the presence or absence of 500 amol sample DNA were analyzed on 1.0% agarose gel electrophoresis. The reaction was conducted at 60°C for 30, 45, 60, 75, 90, 105 and 120 min and each reaction was analyzed separately. Lane M was loaded with a 100 bp DNA marker (100, 200, 300, 400, 500/517, 600, 700, 800, 900, 1000, 1200 and 1517 bp from the bottom). ( B ) Endpoint products of 90-min reactions with all or partial reaction components were analyzed on 1.0% agarose gel electrophoresis. DNA, POL and NICK indicate sample DNA, Vent (exo-) DNA polymerase and Nb.BsmI, respectively. Lane M is a 100 bp DNA marker. ( C ) Endpoint product of a 120-min PG–RCA reaction was digested with either BsmI or BbvCI and analyzed on 1.5% agarose gel electrophoresis. Lane ‘-’ was loaded with the 120 min reaction product before restriction enzyme digestion. Lane M is a 100 bp DNA marker.
    Figure Legend Snippet: Endpoint product analysis of PG–RCA. ( A ) Endpoint products in the presence or absence of 500 amol sample DNA were analyzed on 1.0% agarose gel electrophoresis. The reaction was conducted at 60°C for 30, 45, 60, 75, 90, 105 and 120 min and each reaction was analyzed separately. Lane M was loaded with a 100 bp DNA marker (100, 200, 300, 400, 500/517, 600, 700, 800, 900, 1000, 1200 and 1517 bp from the bottom). ( B ) Endpoint products of 90-min reactions with all or partial reaction components were analyzed on 1.0% agarose gel electrophoresis. DNA, POL and NICK indicate sample DNA, Vent (exo-) DNA polymerase and Nb.BsmI, respectively. Lane M is a 100 bp DNA marker. ( C ) Endpoint product of a 120-min PG–RCA reaction was digested with either BsmI or BbvCI and analyzed on 1.5% agarose gel electrophoresis. Lane ‘-’ was loaded with the 120 min reaction product before restriction enzyme digestion. Lane M is a 100 bp DNA marker.

    Techniques Used: Agarose Gel Electrophoresis, Marker

    5) Product Images from "Determining DNA Supercoiling Enthalpy by Isothermal Titration Calorimetry"

    Article Title: Determining DNA Supercoiling Enthalpy by Isothermal Titration Calorimetry

    Journal: Biochimie

    doi: 10.1016/j.biochi.2012.08.002

    (A) The plasmid map of pXXZ6. Restriction enzyme recognition sites of BamHI, HindIII, and Nt.BbvCI are shown. For supercoiled pXXZ6 used in this study, the supercoiling density was determined to be −0.061. (B) DNA intercalator ethidium bromide
    Figure Legend Snippet: (A) The plasmid map of pXXZ6. Restriction enzyme recognition sites of BamHI, HindIII, and Nt.BbvCI are shown. For supercoiled pXXZ6 used in this study, the supercoiling density was determined to be −0.061. (B) DNA intercalator ethidium bromide

    Techniques Used: Plasmid Preparation

    Related Articles

    Clone Assay:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    Luciferase:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    In Vitro:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    Methylation:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    Construct:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    Purification:

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens
    Article Snippet: .. The PCR products were digested with Bbv CI and Bst XI (New England BioLabs Inc, Ipswich, MA), and gel purified using an AccuPrep™ Gel Purification Kit (Bioneer, Alameda, CA). .. The destination binary plasmid pEarleyGate101 [ ] was obtained from the Arabidopsis Biological Resource Center at Ohio State University (Columbus, OH) and digested with Bbv CI and Bst XI to remove the ccdB gene.

    Generated:

    Article Title: Germline Epigenetic Regulation of KILLIN in Cowden and Cowden-Like Syndromes
    Article Snippet: .. The in vitro methylated constructs used for the luciferase assay were generated by first digesting 90µg of the original PTEN and KILLIN promoter constructs (containing 1 to 1344bp upstream of the PTEN translational start site cloned in either direction) with BglII (NEB) and BbvCI (NEB). .. The insert DNAs, which contain the sequence that is methylated in vivo in CS/CSL patients, were then methylated with CpG SssI methylase (NEB) for 4 hours.

    other:

    Article Title: Sensitive isothermal detection of nucleic-acid sequence by primer generation-rolling circle amplification
    Article Snippet: Vent (exo-) DNA polymerase, Nb.BsmI, BsmI, BbvCI and exonuclease III were purchased from New England Biolabs.

    BAC Assay:

    Article Title: Single molecule linear analysis of DNA in nano-channel labeled with sequence specific fluorescent probes
    Article Snippet: .. BbvCI (0.5 U/µl) (NEB Cat #R063) 1 µl in 1× NEB buffer 2 (Cat #B7002S) in 20 µl volume for 1 h at 37°C and 20 min at 65°C. (ii) Digestion: After nicking reaction, the circular nicked DNA samples (25 ng/µl) (Fosmid G248P8446G6, MCF7 BAC clone 3F5) were digested with NotI (1 U/µl) (NEB Not1.HF Cat #R3189S) in 1× NEB2 buffer in presence of 1× BSA (NEB BSA, Cat #B90015) to make them linear. .. Typically the incubation was performed for 2 h at 37°C followed by 20 min at 65°C. (iii) DNA strand displacement and Flap generation: In this procedure the nicked and cut DNA samples (12.5 ng/µl) were incubated for 30 min at 50°C in 1× NEB thermopol buffer with Vent (exo-) at 0.5 U/µl in presence of 75 nM dNTP mixture.

    Polymerase Chain Reaction:

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens
    Article Snippet: .. The PCR products were digested with Bbv CI and Bst XI (New England BioLabs Inc, Ipswich, MA), and gel purified using an AccuPrep™ Gel Purification Kit (Bioneer, Alameda, CA). .. The destination binary plasmid pEarleyGate101 [ ] was obtained from the Arabidopsis Biological Resource Center at Ohio State University (Columbus, OH) and digested with Bbv CI and Bst XI to remove the ccdB gene.

    Molecular Weight:

    Article Title: Structural diversity of supercoiled DNA
    Article Snippet: .. BbvCI, EcoRV, Nb.BbvCI, NdeI, Nuclease Bal-31, T4 DNA Ligase, low molecular weight DNA ladder and 100 bp DNA ladder were purchased from New England Biolabs (Ipswich, MA). .. Proteinase K was purchased from Roche Molecular Biochemicals (Mannheim, Germany).

    Gel Purification:

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens
    Article Snippet: .. The PCR products were digested with Bbv CI and Bst XI (New England BioLabs Inc, Ipswich, MA), and gel purified using an AccuPrep™ Gel Purification Kit (Bioneer, Alameda, CA). .. The destination binary plasmid pEarleyGate101 [ ] was obtained from the Arabidopsis Biological Resource Center at Ohio State University (Columbus, OH) and digested with Bbv CI and Bst XI to remove the ccdB gene.

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    New England Biolabs bbvci
    Effect of supercoiling on the structure of minicircle DNA. ( a ) Individual 336 bp minicircle topoisomers were isolated and analysed by polyacrylamide gel electrophoresis in the presence of 10 mM CaCl 2 . Mr: 100 bp DNA ladder, L: minicircle linearized by <t>EcoRV,</t> N: minicircle nicked by <t>Nb.BbvCI.</t> ( b ) Projections of cryo-ET subtomograms of hydrated 336 bp DNA minicircles of the Lk =34 topoisomer. ( c ) Commonly observed shapes were open circle, open figure-8, figure-8, racquet, handcuffs, needle, and rod, each of which are shown in orthogonal views. ( d ) Other shapes observed, especially in the more highly supercoiled topoisomers. ( e ) Shape frequency distribution plot for each topoisomer population (n=number of minicircles analysed). A weighted average for each topoisomer, approximating the average degree of compactness, is denoted by the black triangle. The weighted average was calculated by assigning each conformation a value that increased in line with compactness. Open circles were given a value of 1, open figure-8 s a value of 2, figure-8 s as a value of 3, and so on. The relative fraction of each was subsequently used to determine the average degree of compactness. Lk , Δ Lk and superhelical density (σ) for each topoisomer are shown (see Supplementary Note 1 ).
    Bbvci, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bbvci/product/New England Biolabs
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    bbvci - by Bioz Stars, 2020-07
    94/100 stars
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    Effect of supercoiling on the structure of minicircle DNA. ( a ) Individual 336 bp minicircle topoisomers were isolated and analysed by polyacrylamide gel electrophoresis in the presence of 10 mM CaCl 2 . Mr: 100 bp DNA ladder, L: minicircle linearized by EcoRV, N: minicircle nicked by Nb.BbvCI. ( b ) Projections of cryo-ET subtomograms of hydrated 336 bp DNA minicircles of the Lk =34 topoisomer. ( c ) Commonly observed shapes were open circle, open figure-8, figure-8, racquet, handcuffs, needle, and rod, each of which are shown in orthogonal views. ( d ) Other shapes observed, especially in the more highly supercoiled topoisomers. ( e ) Shape frequency distribution plot for each topoisomer population (n=number of minicircles analysed). A weighted average for each topoisomer, approximating the average degree of compactness, is denoted by the black triangle. The weighted average was calculated by assigning each conformation a value that increased in line with compactness. Open circles were given a value of 1, open figure-8 s a value of 2, figure-8 s as a value of 3, and so on. The relative fraction of each was subsequently used to determine the average degree of compactness. Lk , Δ Lk and superhelical density (σ) for each topoisomer are shown (see Supplementary Note 1 ).

    Journal: Nature Communications

    Article Title: Structural diversity of supercoiled DNA

    doi: 10.1038/ncomms9440

    Figure Lengend Snippet: Effect of supercoiling on the structure of minicircle DNA. ( a ) Individual 336 bp minicircle topoisomers were isolated and analysed by polyacrylamide gel electrophoresis in the presence of 10 mM CaCl 2 . Mr: 100 bp DNA ladder, L: minicircle linearized by EcoRV, N: minicircle nicked by Nb.BbvCI. ( b ) Projections of cryo-ET subtomograms of hydrated 336 bp DNA minicircles of the Lk =34 topoisomer. ( c ) Commonly observed shapes were open circle, open figure-8, figure-8, racquet, handcuffs, needle, and rod, each of which are shown in orthogonal views. ( d ) Other shapes observed, especially in the more highly supercoiled topoisomers. ( e ) Shape frequency distribution plot for each topoisomer population (n=number of minicircles analysed). A weighted average for each topoisomer, approximating the average degree of compactness, is denoted by the black triangle. The weighted average was calculated by assigning each conformation a value that increased in line with compactness. Open circles were given a value of 1, open figure-8 s a value of 2, figure-8 s as a value of 3, and so on. The relative fraction of each was subsequently used to determine the average degree of compactness. Lk , Δ Lk and superhelical density (σ) for each topoisomer are shown (see Supplementary Note 1 ).

    Article Snippet: BbvCI, EcoRV, Nb.BbvCI, NdeI, Nuclease Bal-31, T4 DNA Ligase, low molecular weight DNA ladder and 100 bp DNA ladder were purchased from New England Biolabs (Ipswich, MA).

    Techniques: Isolation, Polyacrylamide Gel Electrophoresis

    Preparation of a long branched DNA (eF1). ( A ) pES1 was treated with SphI, PvuII and nicking endonuclease Nb.BbvCI. A partial fragment from pGap1 was prepared by PCR with indicated primers, followed by nicking with Nt.BbvCI. ( B ) Single strands between the nicks introduced in the previous step were dissociated by heating with removal by complementary oligo DNA from the fragment containing a gap structure. ( C ) Resulting DNAs with the gap were annealed to form a branched structure (Materials and methods section). A leftward triangle indicates an EcoR124I site, 5′ CGA TGCTGTA TTC . An open circle indicates 32 P for 5′-end labeling.

    Journal: Nucleic Acids Research

    Article Title: Cleavage of a model DNA replication fork by a Type I restriction endonuclease

    doi: 10.1093/nar/gkp214

    Figure Lengend Snippet: Preparation of a long branched DNA (eF1). ( A ) pES1 was treated with SphI, PvuII and nicking endonuclease Nb.BbvCI. A partial fragment from pGap1 was prepared by PCR with indicated primers, followed by nicking with Nt.BbvCI. ( B ) Single strands between the nicks introduced in the previous step were dissociated by heating with removal by complementary oligo DNA from the fragment containing a gap structure. ( C ) Resulting DNAs with the gap were annealed to form a branched structure (Materials and methods section). A leftward triangle indicates an EcoR124I site, 5′ CGA TGCTGTA TTC . An open circle indicates 32 P for 5′-end labeling.

    Article Snippet: BbvCI, SphI and PvuII (New England Biolabs).

    Techniques: Polymerase Chain Reaction, End Labeling

    Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.

    Journal: Plant Methods

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens

    doi: 10.1186/1746-4811-7-42

    Figure Lengend Snippet: Schematic cloning of a gene into a binary expression vector and selection in Agrobacterium tumefaciens . (A) Key features of pEG101-SacB/R. LB: T-DNA left border; RB: T-DNA right border; Cm R : chloramphenicol resistance gene; attL1 and attL2: recognition sites of the LR clonase; ccdB : the Invitrogen ccdB gene cassette frame B; SacB/SacR : Levansucrase genes along with their native promoter; Bbv CI and Bst XI: restriction enzyme sites for replacing the ccdB gene fragment with the SacB/SacR gene cassette. (B). Compared to the regular LR cloning procedure, the new protocol is more convenient and time-saving.

    Article Snippet: The PCR products were digested with Bbv CI and Bst XI (New England BioLabs Inc, Ipswich, MA), and gel purified using an AccuPrep™ Gel Purification Kit (Bioneer, Alameda, CA).

    Techniques: Clone Assay, Expressing, Plasmid Preparation, Selection

    The destination vector pEG101-SacB/R can be stably propagated in the E. coli strain DH5α . Restriction digestion ( Bst XI and Bbv CI) of plasmid DNAs, isolated from five repeated overnight cultures of E. coli carrying the pEG101-SacB/R destination vector, showed identical restriction patterns. M: 1 kb marker. Lanes 1 through 5 were digested plasmid DNAs of five repeated subcultures.

    Journal: Plant Methods

    Article Title: A novel Gateway®-compatible binary vector allows direct selection of recombinant clones in Agrobacterium tumefaciens

    doi: 10.1186/1746-4811-7-42

    Figure Lengend Snippet: The destination vector pEG101-SacB/R can be stably propagated in the E. coli strain DH5α . Restriction digestion ( Bst XI and Bbv CI) of plasmid DNAs, isolated from five repeated overnight cultures of E. coli carrying the pEG101-SacB/R destination vector, showed identical restriction patterns. M: 1 kb marker. Lanes 1 through 5 were digested plasmid DNAs of five repeated subcultures.

    Article Snippet: The PCR products were digested with Bbv CI and Bst XI (New England BioLabs Inc, Ipswich, MA), and gel purified using an AccuPrep™ Gel Purification Kit (Bioneer, Alameda, CA).

    Techniques: Plasmid Preparation, Stable Transfection, Isolation, Marker

    Endpoint product analysis of PG–RCA. ( A ) Endpoint products in the presence or absence of 500 amol sample DNA were analyzed on 1.0% agarose gel electrophoresis. The reaction was conducted at 60°C for 30, 45, 60, 75, 90, 105 and 120 min and each reaction was analyzed separately. Lane M was loaded with a 100 bp DNA marker (100, 200, 300, 400, 500/517, 600, 700, 800, 900, 1000, 1200 and 1517 bp from the bottom). ( B ) Endpoint products of 90-min reactions with all or partial reaction components were analyzed on 1.0% agarose gel electrophoresis. DNA, POL and NICK indicate sample DNA, Vent (exo-) DNA polymerase and Nb.BsmI, respectively. Lane M is a 100 bp DNA marker. ( C ) Endpoint product of a 120-min PG–RCA reaction was digested with either BsmI or BbvCI and analyzed on 1.5% agarose gel electrophoresis. Lane ‘-’ was loaded with the 120 min reaction product before restriction enzyme digestion. Lane M is a 100 bp DNA marker.

    Journal: Nucleic Acids Research

    Article Title: Sensitive isothermal detection of nucleic-acid sequence by primer generation-rolling circle amplification

    doi: 10.1093/nar/gkn1014

    Figure Lengend Snippet: Endpoint product analysis of PG–RCA. ( A ) Endpoint products in the presence or absence of 500 amol sample DNA were analyzed on 1.0% agarose gel electrophoresis. The reaction was conducted at 60°C for 30, 45, 60, 75, 90, 105 and 120 min and each reaction was analyzed separately. Lane M was loaded with a 100 bp DNA marker (100, 200, 300, 400, 500/517, 600, 700, 800, 900, 1000, 1200 and 1517 bp from the bottom). ( B ) Endpoint products of 90-min reactions with all or partial reaction components were analyzed on 1.0% agarose gel electrophoresis. DNA, POL and NICK indicate sample DNA, Vent (exo-) DNA polymerase and Nb.BsmI, respectively. Lane M is a 100 bp DNA marker. ( C ) Endpoint product of a 120-min PG–RCA reaction was digested with either BsmI or BbvCI and analyzed on 1.5% agarose gel electrophoresis. Lane ‘-’ was loaded with the 120 min reaction product before restriction enzyme digestion. Lane M is a 100 bp DNA marker.

    Article Snippet: Vent (exo-) DNA polymerase, Nb.BsmI, BsmI, BbvCI and exonuclease III were purchased from New England Biolabs.

    Techniques: Agarose Gel Electrophoresis, Marker