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  • 95
    New England Biolabs bsa i
    Restriction fragment length polymorphism analysis of <t>23S</t> rRNA amplicons: ( A ) digestion with <t>Bsa</t> I and ( B ) digestion with Bbs I. The A2144G mutations are observed in lanes 1 to 2, but not in lanes 3 to 5. Note that the A2143G mutation detected by digestion with Bbs I was not detected in any of the strains studied. Lanes 3 to 5 reveal the T2183C mutation, as assessed by DNA sequencing. Lane M, 100 bp DNA size markers (indicated to the left of the gels in base pairs); lane C, H. pylori ATCC 43504; lane 1 to 5, clarithromycin-resistant H. pylori strains.
    Bsa I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 443 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    bsa i - by Bioz Stars, 2020-02
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    90
    Thermo Fisher eco31i
    Construction of MIDGE vectors . (A) POMC exon 2-3 cDNA, encoding the signal peptide, adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormone (MSH) and β-endorphin (END), was spliced into the pMOK plasmid between the SacI and KpnI restriction sites. The pMOK plasmid contains a kanamycin resistance gene, a cytomegalovirus promoter (PCMV), a chimeric intron and simian virus (SV) 40 polyadenylation (pA) site. The POMC-MIDGE-NLS was derived from the pMOK-POMC plasmid by cutting it with <t>Eco31I,</t> ligation with hairpin ODNs at both ends, and coupling of nuclear localization sequence (NLS) to one of the hairpin ODN. The other vectors were prepared analogously by modifying POMC gene as follows: (B) END sequence was removed to obtain control MIDGE-NLS; (C) ACTH and MSH fragments were removed while END sequence was preserved in its original place to obtain 1xEND-MIDGE-NLS encoding 1 copy of END; (D) ACTH fragment was removed, original END sequence was preserved and MSH fragment was replaced with an additional sequence of END to obtain 2xEND-MIDGE-NLS encoding 2 copies of END; (E) original END sequence was preserved and the ACTH and MSH fragments were replaced with additional END sequences to obtain 3xEND-MIDGE-NLS encoding 3 copies of END.
    Eco31i, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 76 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs bsa i hf
    Construction strategy of the low-copy pMKP ccd B reverse genetic vector to clone unstable HA segments of IAV. The resulting vector is a hybrid of pSMART-LC-Kan and the Pol-I/ ccd B/Pol-II cassette of pMP ccd B with either AarI or <t>Bsa</t> I or BsmB I sites for restriction/cloning of the desired insert. Briefly, the sequence representing the Pol-I/ ccd B/Pol-II cassette excised from pMP ccd B by digestion with BsrB I were blunt end phosphorylated, and ligated into the EcoR V linearized pSMART-LC-Kan plasmid to generate pMKP ccd B. The pMKP ccd B was suitable to clone and maintain the genetically unstable RT-PCR product of H9N2/SA- and H5N1/VSVRI-HA. Transcriptional terminator (T); Repressor of Primer (ROP); low copy origin of replication (Ori); kanamycin resistance gene (Kanr), ampicillin resistance gene (Ampr).
    Bsa I Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher bsai
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs bsai hfv2
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Hfv2, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    LifeSensors bsai site
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Site, supplied by LifeSensors, used in various techniques. Bioz Stars score: 83/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore 2g12lc bsai fw 2g12lc bsai rv
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    2g12lc Bsai Fw 2g12lc Bsai Rv, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher bsai fd
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Fd, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher fastdigest bsai
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Fastdigest Bsai, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher bsai mboii
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Mboii, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Addgene inc bsai digested pdr274
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Digested Pdr274, supplied by Addgene inc, used in various techniques. Bioz Stars score: 85/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    5 PRIME bsai
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 99/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Addgene inc bsai digested prb1017
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    Bsai Digested Prb1017, supplied by Addgene inc, used in various techniques. Bioz Stars score: 91/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Addgene inc u6 bsai sgrna
    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With <t>BsaI</t> and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.
    U6 Bsai Sgrna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 80/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    New England Biolabs bsai enzyme
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 92/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    New England Biolabs bsai reases
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Reases, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 84/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Addgene inc bsai cut pu6 universal
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Cut Pu6 Universal, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher bsai cloning sites
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Cloning Sites, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher bsai linearized pe sumo
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Linearized Pe Sumo, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher bsai hf
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Hf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Thermo Fisher bsai site
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Site, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 82/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    New England Biolabs bsai buffer
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Addgene inc bsai digested dr274 vector
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Digested Dr274 Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 80/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 369 pentrattl4attr1 bsai vector
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
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    GenScript bsai restriction sequences
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Restriction Sequences, supplied by GenScript, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc puckanr mu bsai
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Puckanr Mu Bsai, supplied by Addgene inc, used in various techniques. Bioz Stars score: 84/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GenScript bsai recognition sites
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Bsai Recognition Sites, supplied by GenScript, used in various techniques. Bioz Stars score: 83/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Addgene inc bsai digested prgeb31 vectors
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
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    New England Biolabs bsai hf
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
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    Genewiz terminator cassette l1 ttpi1 aggtt adapter bsai bsai aatcg adapter pgit l2
    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by <t>BsaI</t> restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb <t>DNA</t> Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.
    Terminator Cassette L1 Ttpi1 Aggtt Adapter Bsai Bsai Aatcg Adapter Pgit L2, supplied by Genewiz, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Restriction fragment length polymorphism analysis of 23S rRNA amplicons: ( A ) digestion with Bsa I and ( B ) digestion with Bbs I. The A2144G mutations are observed in lanes 1 to 2, but not in lanes 3 to 5. Note that the A2143G mutation detected by digestion with Bbs I was not detected in any of the strains studied. Lanes 3 to 5 reveal the T2183C mutation, as assessed by DNA sequencing. Lane M, 100 bp DNA size markers (indicated to the left of the gels in base pairs); lane C, H. pylori ATCC 43504; lane 1 to 5, clarithromycin-resistant H. pylori strains.

    Journal: Journal of Korean Medical Science

    Article Title: Clarithromycin-Based Standard Triple Therapy Can Still Be Effective for Helicobacter pylori Eradication in Some Parts of the Korea

    doi: 10.3346/jkms.2014.29.9.1240

    Figure Lengend Snippet: Restriction fragment length polymorphism analysis of 23S rRNA amplicons: ( A ) digestion with Bsa I and ( B ) digestion with Bbs I. The A2144G mutations are observed in lanes 1 to 2, but not in lanes 3 to 5. Note that the A2143G mutation detected by digestion with Bbs I was not detected in any of the strains studied. Lanes 3 to 5 reveal the T2183C mutation, as assessed by DNA sequencing. Lane M, 100 bp DNA size markers (indicated to the left of the gels in base pairs); lane C, H. pylori ATCC 43504; lane 1 to 5, clarithromycin-resistant H. pylori strains.

    Article Snippet: Amplicons (424 bp each) of the 23S rRNA gene were digested with either Bsa I (New England BioLabs, Beverly, MA, USA) for 14 hr at 50℃ or Bbs I (New England BioLabs) for 14 hr at 37℃ to detect the A2144G and A2143G mutations, respectively ( ).

    Techniques: Mutagenesis, DNA Sequencing

    Construction of MIDGE vectors . (A) POMC exon 2-3 cDNA, encoding the signal peptide, adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormone (MSH) and β-endorphin (END), was spliced into the pMOK plasmid between the SacI and KpnI restriction sites. The pMOK plasmid contains a kanamycin resistance gene, a cytomegalovirus promoter (PCMV), a chimeric intron and simian virus (SV) 40 polyadenylation (pA) site. The POMC-MIDGE-NLS was derived from the pMOK-POMC plasmid by cutting it with Eco31I, ligation with hairpin ODNs at both ends, and coupling of nuclear localization sequence (NLS) to one of the hairpin ODN. The other vectors were prepared analogously by modifying POMC gene as follows: (B) END sequence was removed to obtain control MIDGE-NLS; (C) ACTH and MSH fragments were removed while END sequence was preserved in its original place to obtain 1xEND-MIDGE-NLS encoding 1 copy of END; (D) ACTH fragment was removed, original END sequence was preserved and MSH fragment was replaced with an additional sequence of END to obtain 2xEND-MIDGE-NLS encoding 2 copies of END; (E) original END sequence was preserved and the ACTH and MSH fragments were replaced with additional END sequences to obtain 3xEND-MIDGE-NLS encoding 3 copies of END.

    Journal: Molecular Pain

    Article Title: Peripheral non-viral MIDGE vector-driven delivery of ?-endorphin in inflammatory pain

    doi: 10.1186/1744-8069-5-72

    Figure Lengend Snippet: Construction of MIDGE vectors . (A) POMC exon 2-3 cDNA, encoding the signal peptide, adrenocorticotropic hormone (ACTH), melanocyte-stimulating hormone (MSH) and β-endorphin (END), was spliced into the pMOK plasmid between the SacI and KpnI restriction sites. The pMOK plasmid contains a kanamycin resistance gene, a cytomegalovirus promoter (PCMV), a chimeric intron and simian virus (SV) 40 polyadenylation (pA) site. The POMC-MIDGE-NLS was derived from the pMOK-POMC plasmid by cutting it with Eco31I, ligation with hairpin ODNs at both ends, and coupling of nuclear localization sequence (NLS) to one of the hairpin ODN. The other vectors were prepared analogously by modifying POMC gene as follows: (B) END sequence was removed to obtain control MIDGE-NLS; (C) ACTH and MSH fragments were removed while END sequence was preserved in its original place to obtain 1xEND-MIDGE-NLS encoding 1 copy of END; (D) ACTH fragment was removed, original END sequence was preserved and MSH fragment was replaced with an additional sequence of END to obtain 2xEND-MIDGE-NLS encoding 2 copies of END; (E) original END sequence was preserved and the ACTH and MSH fragments were replaced with additional END sequences to obtain 3xEND-MIDGE-NLS encoding 3 copies of END.

    Article Snippet: The product was cut with Eco31I and SacI, ligated and cloned into the pMOK-POMC1xEND plasmid after digestion with SacI and BpiI (Fermentas).

    Techniques: Plasmid Preparation, Derivative Assay, Ligation, Sequencing

    Construction strategy of the low-copy pMKP ccd B reverse genetic vector to clone unstable HA segments of IAV. The resulting vector is a hybrid of pSMART-LC-Kan and the Pol-I/ ccd B/Pol-II cassette of pMP ccd B with either AarI or Bsa I or BsmB I sites for restriction/cloning of the desired insert. Briefly, the sequence representing the Pol-I/ ccd B/Pol-II cassette excised from pMP ccd B by digestion with BsrB I were blunt end phosphorylated, and ligated into the EcoR V linearized pSMART-LC-Kan plasmid to generate pMKP ccd B. The pMKP ccd B was suitable to clone and maintain the genetically unstable RT-PCR product of H9N2/SA- and H5N1/VSVRI-HA. Transcriptional terminator (T); Repressor of Primer (ROP); low copy origin of replication (Ori); kanamycin resistance gene (Kanr), ampicillin resistance gene (Ampr).

    Journal: PLoS ONE

    Article Title: Efficient Generation of Recombinant Influenza A Viruses Employing a New Approach to Overcome the Genetic Instability of HA Segments

    doi: 10.1371/journal.pone.0116917

    Figure Lengend Snippet: Construction strategy of the low-copy pMKP ccd B reverse genetic vector to clone unstable HA segments of IAV. The resulting vector is a hybrid of pSMART-LC-Kan and the Pol-I/ ccd B/Pol-II cassette of pMP ccd B with either AarI or Bsa I or BsmB I sites for restriction/cloning of the desired insert. Briefly, the sequence representing the Pol-I/ ccd B/Pol-II cassette excised from pMP ccd B by digestion with BsrB I were blunt end phosphorylated, and ligated into the EcoR V linearized pSMART-LC-Kan plasmid to generate pMKP ccd B. The pMKP ccd B was suitable to clone and maintain the genetically unstable RT-PCR product of H9N2/SA- and H5N1/VSVRI-HA. Transcriptional terminator (T); Repressor of Primer (ROP); low copy origin of replication (Ori); kanamycin resistance gene (Kanr), ampicillin resistance gene (Ampr).

    Article Snippet: The positive and correct vector/insert constructs were digested with Bsa I-HF (NEB, Germany) for 15 min at 37°C.

    Techniques: Plasmid Preparation, Clone Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction

    Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With BsaI and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Detection of 23SrRNA Mutations Strongly Related to Clarithromycin Resistance in Helicobacter pylori Strains Isolated From Patients in the North of Iran

    doi: 10.5812/jjm.29694

    Figure Lengend Snippet: Gel Electrophoresis of Digested 429 bp Polymerase Chain Reaction Product of 23S rRNA Gene With BsaI and MboII Enzymes Lane 1, DNA ladder (100 bp); Lane 2, clarithromycin resistant strain digested with BsaI at two sites; Lane 3 and 4, wild types of H. pylori (susceptible to clarithromycin) digested with BsaI, with one digestion site; Lane 5, wild type (susceptible to clarithromycin) treated with MboII with no digestion site.

    Article Snippet: To detect the A-G point mutations at positions 2142 and 2143, the PCR products were digested with the MboII and BsaI (MBI Fermentas, Lithuania) restriction enzymes, respectively, according to the manufacturer’s instructions, and analyzed on 2% agarose gel containing Sybrsafe.

    Techniques: Nucleic Acid Electrophoresis, Polymerase Chain Reaction

    Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by BsaI restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb DNA Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.

    Journal: PLoS ONE

    Article Title: Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes

    doi: 10.1371/journal.pone.0005553

    Figure Lengend Snippet: Assembly of a GFP construct from 10 plasmids. (A) Construct maps. Input modules contain a core region C flanked by BsaI restriction sites in opposite orientation composed of a recognition site (B, ggtctcn, B , ngagacc) and a 4 nucleotide cleavage site (boxes flanking the core region). pX-LacZ, acceptor vector. pGFPi, resulting construct. Restriction sites for AvrII and XmaI are shown as white arrows. (B) Ethidium bromide-stained gel with products obtained by restriction-ligation of the 9 input module plasmids. M: GeneRuler 1kb DNA Ladder Plus from Fermentas. Restriction-ligation was performed at 37°C for 3 (lane 3h) or 6 hours (lane 6h) or with 25 cycles (2 min 37°C+5 min 16°C, lane 25) or 50 cycles (lane 50), and without BsaI enzyme (lane nb). The arrow indicates the 1.17 kb linear assembled GFP gene product. (C) Ethidium bromide-stained gels of 72 minipreps digested with XmaI and AvrII (expected fragment sizes: 4.6 kb, 945 and 555 bp), obtained from restriction-ligations performed for 6 h 37°C (6 h), for 25 or 50 cycles (25×/50×), with normal ligase (nl) or high concentration ligase (hcl). Numbers indicate minipreps with an incorrect restriction pattern, and stars indicate constructs that consist of dimers (same restriction pattern as monomers). V, vector pX-lacZ.

    Article Snippet: A restriction-ligation was set up by adding into a single tube 50 ng of each of the 27 trypsinogen fragment constructs , 50 ng of vector, 10 units of BsaI enzyme (NEB) and 3 units of T4 DNA ligase (Promega) in a total volume of 15 microliters in ligation buffer (Promega).

    Techniques: Construct, Plasmid Preparation, Staining, Ligation, Concentration Assay

    DNA shuffling strategy. (A) Two DNA ends terminated by the same 4 nucleotides (sequence f, composed of nucleotides 1234, complementary nucleotides noted in italics) flanked by a BsaI recognition sequence, B, form two complementary DNA overhangs after digestion with BsaI. (B) For shuffling, genes of interest are aligned, and recombination points consisting of 4 nucleotide sequences (f1 to fn+1) are defined on conserved sequences. Module fragments (core sequence, C1 to Cn, plus flanking 4 nucleotide sequences) are amplified by PCR and cloned in an intermediate cloning vector. Module fragment plasmids and the acceptor vector are assembled in one restriction-ligation with BsaI and ligase. S1 and S2, two different selectable markers. Z, lacZ alpha gene fragment.

    Journal: PLoS ONE

    Article Title: Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes

    doi: 10.1371/journal.pone.0005553

    Figure Lengend Snippet: DNA shuffling strategy. (A) Two DNA ends terminated by the same 4 nucleotides (sequence f, composed of nucleotides 1234, complementary nucleotides noted in italics) flanked by a BsaI recognition sequence, B, form two complementary DNA overhangs after digestion with BsaI. (B) For shuffling, genes of interest are aligned, and recombination points consisting of 4 nucleotide sequences (f1 to fn+1) are defined on conserved sequences. Module fragments (core sequence, C1 to Cn, plus flanking 4 nucleotide sequences) are amplified by PCR and cloned in an intermediate cloning vector. Module fragment plasmids and the acceptor vector are assembled in one restriction-ligation with BsaI and ligase. S1 and S2, two different selectable markers. Z, lacZ alpha gene fragment.

    Article Snippet: A restriction-ligation was set up by adding into a single tube 50 ng of each of the 27 trypsinogen fragment constructs , 50 ng of vector, 10 units of BsaI enzyme (NEB) and 3 units of T4 DNA ligase (Promega) in a total volume of 15 microliters in ligation buffer (Promega).

    Techniques: Sequencing, Amplification, Polymerase Chain Reaction, Clone Assay, Plasmid Preparation, Ligation

    Shuffling of trypsinogen. (A) Alignment of the aminoacid sequence of bovine cationic trypsinogen (BC), bovine anionic trypsinogen (BA) and human cationic trypsinogen (HC). Nucleotide sequence of the chosen recombination sites is shown. (B) Map of the 27 trypsinogen module plasmids, the acceptor vector, and of an example of one of the resulting shuffled construct obtained. B, BsaI restriction site. S, K: spectinomycin and kanamycin resistance genes. RB/LB, T-DNA right and left borders. AttB, Phage C31 recombination site, N tobamoviral 3′ non-translated region, T, Nos terminator. (C) Ethidium bromide-stained gels of 28 minipreps prepared from single colonies (1 to 24) or from 4 libraries (L1–4, approximately 700 clones in each) digested with XmaI (incorrect pattern 1, 2, 7 and 17).

    Journal: PLoS ONE

    Article Title: Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes

    doi: 10.1371/journal.pone.0005553

    Figure Lengend Snippet: Shuffling of trypsinogen. (A) Alignment of the aminoacid sequence of bovine cationic trypsinogen (BC), bovine anionic trypsinogen (BA) and human cationic trypsinogen (HC). Nucleotide sequence of the chosen recombination sites is shown. (B) Map of the 27 trypsinogen module plasmids, the acceptor vector, and of an example of one of the resulting shuffled construct obtained. B, BsaI restriction site. S, K: spectinomycin and kanamycin resistance genes. RB/LB, T-DNA right and left borders. AttB, Phage C31 recombination site, N tobamoviral 3′ non-translated region, T, Nos terminator. (C) Ethidium bromide-stained gels of 28 minipreps prepared from single colonies (1 to 24) or from 4 libraries (L1–4, approximately 700 clones in each) digested with XmaI (incorrect pattern 1, 2, 7 and 17).

    Article Snippet: A restriction-ligation was set up by adding into a single tube 50 ng of each of the 27 trypsinogen fragment constructs , 50 ng of vector, 10 units of BsaI enzyme (NEB) and 3 units of T4 DNA ligase (Promega) in a total volume of 15 microliters in ligation buffer (Promega).

    Techniques: Sequencing, Plasmid Preparation, Construct, Staining, Clone Assay