bsrgi New England Biolabs Search Results


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  • 90
    New England Biolabs bsrg i
    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using <t>Hha</t> I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and <t>BsrG</t> I endonucleases.
    Bsrg I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    New England Biolabs bsrgi hf
    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using <t>Hha</t> I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and <t>BsrG</t> I endonucleases.
    Bsrgi Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 84/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    New England Biolabs bsrg1
    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using <t>Hha</t> I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and <t>BsrG</t> I endonucleases.
    Bsrg1, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    New England Biolabs nsp i
    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using <t>Hha</t> I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and <t>BsrG</t> I endonucleases.
    Nsp I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 87/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs bsrg i restriction enzymes
    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using <t>Hha</t> I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and <t>BsrG</t> I endonucleases.
    Bsrg I Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    New England Biolabs enzyme bsrg i
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Enzyme Bsrg I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 78/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    New England Biolabs bsrgl
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Bsrgl, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 79/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    New England Biolabs bsrgi enzyme
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Bsrgi Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    New England Biolabs acc65i
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Acc65i, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs eagi
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Eagi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 313 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs bsm bi
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    Bsm Bi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 164 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs t5 exonuclease
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    T5 Exonuclease, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 488 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs t4 pnk enzyme
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    T4 Pnk Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs t4 pnk
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    T4 Pnk, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3038 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs t4 polynucleotide kinase
    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with <t>BsrG</t> 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.
    T4 Polynucleotide Kinase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 22375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs sau96i
    RFLP profile of the URA5 genes from Cryptococcus sp obtained by double-digestion with HhaI and <t>Sau96I.</t> Column 1: Molecular maker (100 bp). Columns 2 to 7: isolated 2, 4, 5, 6, 7 and 8 (VNI), respectively; column 8: isolated 12 (VGI); Columns 9 to 13: isolates 13, 18, 22, 24 and 25 (VNI), respectively.
    Sau96i, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs rnase inhibitor
    RFLP profile of the URA5 genes from Cryptococcus sp obtained by double-digestion with HhaI and <t>Sau96I.</t> Column 1: Molecular maker (100 bp). Columns 2 to 7: isolated 2, 4, 5, 6, 7 and 8 (VNI), respectively; column 8: isolated 12 (VGI); Columns 9 to 13: isolates 13, 18, 22, 24 and 25 (VNI), respectively.
    Rnase Inhibitor, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1529 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs restriction endonuclease bsrg1
    RFLP profile of the URA5 genes from Cryptococcus sp obtained by double-digestion with HhaI and <t>Sau96I.</t> Column 1: Molecular maker (100 bp). Columns 2 to 7: isolated 2, 4, 5, 6, 7 and 8 (VNI), respectively; column 8: isolated 12 (VGI); Columns 9 to 13: isolates 13, 18, 22, 24 and 25 (VNI), respectively.
    Restriction Endonuclease Bsrg1, supplied by New England Biolabs, 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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    99
    New England Biolabs phi29 dna polymerase
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    Phi29 Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 558 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    74
    New England Biolabs restriction enzymes
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 74/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs t4 dna ligase
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    T4 Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 36171 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs restriction endonuclease dpni
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    Restriction Endonuclease Dpni, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs m mulv reverse transcriptase
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    M Mulv Reverse Transcriptase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3431 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs amv reverse transcriptase
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    Amv Reverse Transcriptase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 450 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    New England Biolabs dna e coli polymerase i
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    Dna E Coli Polymerase I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 79/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs e coli dna ligase
    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 <t>DNA</t> as template and <t>phi29</t> DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.
    E Coli Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 384 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    e coli dna ligase - by Bioz Stars, 2020-01
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    New England Biolabs t7 endonuclease i
    Delivery of Cas9:sgRNA, Cas9 D10A nickase, and dCas9-VP64 transcriptional activators to cultured human cells. ( a ) Green entries: U2OS EGFP reporter cells were treated with 100 nM of the Cas9 protein variant shown, 0.8 µL of the cationic lipid shown, and either 50 nM of the sgRNA shown for Cas9 protein treatment, or 125 nM of the sgRNA shown for (+36)dGFP-NLS-Cas9 and (−30)dGFP-NLS-Cas9 treatment. The fraction of cells lacking EGFP expression was measured by flow cytometry. Blue entries: plasmid DNA transfection of 750 ng Cas9 and 250 ng sgRNA expression plasmids using 0.8 µL Lipofectamine 2000. ( b ) T7 endonuclease I (T7EI) assay to measure modification of EGFP from no treatment (lane 1), treatment with EGFP-targeting sgRNA alone (lane 2), Cas9 protein alone (lane 3), Cas9 protein + VEGF-targeting sgRNA + RNAiMAX (lane 4), DNA transfection of plasmids expressing Cas9 and EGFP-targeting sgRNA (lane 5), or Cas9 protein + EGFP-targeting sgRNA + RNAiMAX (lane 6). ( c ) T7EI assay of simultaneous genome modification at  EGFP  and three endogenous genes in U2OS cells 48 hours after a single treatment of 100 nM Cas9 protein, 25 nM of each of the four sgRNAs shown (100 nM total sgRNA), and 0.8 µL RNAiMAX. ( d ) Delivery of Cas9 D10A nickase and pairs of sgRNAs either by plasmid transfection or by RNAiMAX-mediated protein:sgRNA complex delivery under conditions described in ( a ) with 50 nM  EGFP -disrupting sgRNAs (25 nM each) for protein delivery, and 250 ng sgRNA-expressing plasmids (125 ng each) for DNA delivery.  EGFP -disrupting sgRNAs g1 + g5, or g3 + g7, are expected to result in gene disruption, while g5 + g7 target the same strand and are expected to be non-functional. ( e ) Delivery of dCas9-VP64 transcriptional activators that target  NTF3  either by DNA transfection or RNAiMAX-mediated protein delivery. Error bars reflect s.d. from six biological replicates performed on different days.
    T7 Endonuclease I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2593 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    t7 endonuclease i - by Bioz Stars, 2020-01
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    Image Search Results


    URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using Hha I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and BsrG I endonucleases.

    Journal: Emerging Infectious Diseases

    Article Title: Spread of Cryptococcus gattii in British Columbia, Canada, and Detection in the Pacific Northwest, USA

    doi: 10.3201/eid1301.060827

    Figure Lengend Snippet: URA5–restriction fragment length polymorphism (RFLP) profiles for selected human, animal, and environmental Cryptococcus gattii isolates. A) URA5-RFLP to determine the molecular type using Hha I and Sau96 I endonucleases ( 14 ). B) URA5-RFLP to confirm molecular type and determine VGII subtype, using Hha I, Dde I, and BsrG I endonucleases.

    Article Snippet: The URA5 gene was amplified as previously described ( ) and then completely digested at 37°C in a 20-μL reaction containing 1× NEB2 buffer, 1× bovine serum albumin, and 4 U each of Hha I, Dde I, and BsrG I (New England Biolabs, Inc., Ipswich, MA, USA).

    Techniques:

    Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with BsrG 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.

    Journal: BMC Research Notes

    Article Title: An improved method for RNA isolation and cDNA library construction from immature seeds of Jatropha curcas L

    doi: 10.1186/1756-0500-3-126

    Figure Lengend Snippet: Evaluation of jatropha seed cDNA library . Poly(A) + RNA was purified from total RNA extracted with method II and used for cDNA library construction using CloneMiner™ cDNA Library Construction Kit (Invitrogen). Plasmid DNA of 20 positive clones was digested with BsrG 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA. (Lane 1) 1-kb DNA marker (New England Biolabs); (lane 2) vector pDONR™ 222 (Invitrogen); (Lanes 3-22) randomly picked cDNA clones. Band at size 2.5 Kb is the vector backbone. cDNA insert of the clone in lane 3 has similar size as that of vector backbone, which did not separate in this gel electrophoresis.

    Article Snippet: The mini-prepared plasmids were digested with BsrG 1 enzyme (New England Biolabs) and electrophoresed on 1% agarose gel to determine average insert size of cDNA.

    Techniques: cDNA Library Assay, Purification, Plasmid Preparation, Clone Assay, Agarose Gel Electrophoresis, Marker, Nucleic Acid Electrophoresis

    RFLP profile of the URA5 genes from Cryptococcus sp obtained by double-digestion with HhaI and Sau96I. Column 1: Molecular maker (100 bp). Columns 2 to 7: isolated 2, 4, 5, 6, 7 and 8 (VNI), respectively; column 8: isolated 12 (VGI); Columns 9 to 13: isolates 13, 18, 22, 24 and 25 (VNI), respectively.

    Journal: Revista do Instituto de Medicina Tropical de São Paulo

    Article Title: The epidemiology of cryptococcosis and the characterization of Cryptococcus neoformans isolated in a Brazilian University Hospital

    doi: 10.1590/S1678-9946201759013

    Figure Lengend Snippet: RFLP profile of the URA5 genes from Cryptococcus sp obtained by double-digestion with HhaI and Sau96I. Column 1: Molecular maker (100 bp). Columns 2 to 7: isolated 2, 4, 5, 6, 7 and 8 (VNI), respectively; column 8: isolated 12 (VGI); Columns 9 to 13: isolates 13, 18, 22, 24 and 25 (VNI), respectively.

    Article Snippet: Thirty microliters of the reaction amplicon were doubly digested with Sau96I (10 U/µL) and HhaI (20 U/µL) (New England Biolabs, Uniscience, SP, Brazil), and incubated at 37 °C for 3 hours.

    Techniques: Isolation

    Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 DNA as template and phi29 DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.

    Journal: Nucleic Acids Research

    Article Title: Improvements of rolling circle amplification (RCA) efficiency and accuracy using Thermus thermophilus SSB mutant protein

    doi: 10.1093/nar/gkl350

    Figure Lengend Snippet: Effect of Tth SSB-255 protein on the efficiency and specificity of RCA. ( a ) Top: RCAs were performed in the absence of the SSB proteins using pUC19 DNA as template and phi29 DNA polymerase for the indicated reaction times. Bottom: Signals of spot hybridization of the same samples. ( b ) Top and Bottom: same as (a), except for the absence of template DNA. ( c ) Top and Bottom: same as (a), except for the presence of the Tth SSB protein (3.0 µg/20 µl reaction volume). ( d ) Top and Bottom: same as (c), except for the absence of template DNA. ( e ) Top and Bottom: same as (a), except for the presence of the Tth SSB-255 protein (3.0 µg/20 µl reaction volume). ( f ) Top and Bottom: same as (e), except for the absence of template DNA.

    Article Snippet: The denatured DNA was amplified using 0.3 µl Phi29 DNA polymerase (10 U/µl, New England Biolabs) complemented with 2 µl of 10× Phi29 DNA polymerase buffer, 0.2 µl of 100× BSA, 3.2 µl of 2.5 mM dNTP and 1 µl of 20% DMSO (Sigma Aldrich) in a volume of 20 µl at 30°C for ∼24 h. The phi29 DNA polymerase was inactivated at 65°C for 10 min and the amplification product was purified using a spin-column (Sephadex G-50, Amersham Biosciences) to eliminate the un-reacted primers.

    Techniques: Hybridization

    Effect of Tth SSB-255 protein on RCA assays. ( a ) RCAs were carried out in the absence or presence of the indicated SSB proteins using pUC19 DNA as a template and phi29 DNA polymerase. ( b ) Same as (a), except for using linearized ( Eco RI) pUC19 DNA as the template. ( c ) Same as (a), except that the amplifications were carried out in the absence of template DNA. Lane M: molecular weight markers (100 and 12 kb).

    Journal: Nucleic Acids Research

    Article Title: Improvements of rolling circle amplification (RCA) efficiency and accuracy using Thermus thermophilus SSB mutant protein

    doi: 10.1093/nar/gkl350

    Figure Lengend Snippet: Effect of Tth SSB-255 protein on RCA assays. ( a ) RCAs were carried out in the absence or presence of the indicated SSB proteins using pUC19 DNA as a template and phi29 DNA polymerase. ( b ) Same as (a), except for using linearized ( Eco RI) pUC19 DNA as the template. ( c ) Same as (a), except that the amplifications were carried out in the absence of template DNA. Lane M: molecular weight markers (100 and 12 kb).

    Article Snippet: The denatured DNA was amplified using 0.3 µl Phi29 DNA polymerase (10 U/µl, New England Biolabs) complemented with 2 µl of 10× Phi29 DNA polymerase buffer, 0.2 µl of 100× BSA, 3.2 µl of 2.5 mM dNTP and 1 µl of 20% DMSO (Sigma Aldrich) in a volume of 20 µl at 30°C for ∼24 h. The phi29 DNA polymerase was inactivated at 65°C for 10 min and the amplification product was purified using a spin-column (Sephadex G-50, Amersham Biosciences) to eliminate the un-reacted primers.

    Techniques: Molecular Weight

    Delivery of Cas9:sgRNA, Cas9 D10A nickase, and dCas9-VP64 transcriptional activators to cultured human cells. ( a ) Green entries: U2OS EGFP reporter cells were treated with 100 nM of the Cas9 protein variant shown, 0.8 µL of the cationic lipid shown, and either 50 nM of the sgRNA shown for Cas9 protein treatment, or 125 nM of the sgRNA shown for (+36)dGFP-NLS-Cas9 and (−30)dGFP-NLS-Cas9 treatment. The fraction of cells lacking EGFP expression was measured by flow cytometry. Blue entries: plasmid DNA transfection of 750 ng Cas9 and 250 ng sgRNA expression plasmids using 0.8 µL Lipofectamine 2000. ( b ) T7 endonuclease I (T7EI) assay to measure modification of EGFP from no treatment (lane 1), treatment with EGFP-targeting sgRNA alone (lane 2), Cas9 protein alone (lane 3), Cas9 protein + VEGF-targeting sgRNA + RNAiMAX (lane 4), DNA transfection of plasmids expressing Cas9 and EGFP-targeting sgRNA (lane 5), or Cas9 protein + EGFP-targeting sgRNA + RNAiMAX (lane 6). ( c ) T7EI assay of simultaneous genome modification at  EGFP  and three endogenous genes in U2OS cells 48 hours after a single treatment of 100 nM Cas9 protein, 25 nM of each of the four sgRNAs shown (100 nM total sgRNA), and 0.8 µL RNAiMAX. ( d ) Delivery of Cas9 D10A nickase and pairs of sgRNAs either by plasmid transfection or by RNAiMAX-mediated protein:sgRNA complex delivery under conditions described in ( a ) with 50 nM  EGFP -disrupting sgRNAs (25 nM each) for protein delivery, and 250 ng sgRNA-expressing plasmids (125 ng each) for DNA delivery.  EGFP -disrupting sgRNAs g1 + g5, or g3 + g7, are expected to result in gene disruption, while g5 + g7 target the same strand and are expected to be non-functional. ( e ) Delivery of dCas9-VP64 transcriptional activators that target  NTF3  either by DNA transfection or RNAiMAX-mediated protein delivery. Error bars reflect s.d. from six biological replicates performed on different days.

    Journal: Nature biotechnology

    Article Title: Efficient Delivery of Genome-Editing Proteins In Vitro and In Vivo

    doi: 10.1038/nbt.3081

    Figure Lengend Snippet: Delivery of Cas9:sgRNA, Cas9 D10A nickase, and dCas9-VP64 transcriptional activators to cultured human cells. ( a ) Green entries: U2OS EGFP reporter cells were treated with 100 nM of the Cas9 protein variant shown, 0.8 µL of the cationic lipid shown, and either 50 nM of the sgRNA shown for Cas9 protein treatment, or 125 nM of the sgRNA shown for (+36)dGFP-NLS-Cas9 and (−30)dGFP-NLS-Cas9 treatment. The fraction of cells lacking EGFP expression was measured by flow cytometry. Blue entries: plasmid DNA transfection of 750 ng Cas9 and 250 ng sgRNA expression plasmids using 0.8 µL Lipofectamine 2000. ( b ) T7 endonuclease I (T7EI) assay to measure modification of EGFP from no treatment (lane 1), treatment with EGFP-targeting sgRNA alone (lane 2), Cas9 protein alone (lane 3), Cas9 protein + VEGF-targeting sgRNA + RNAiMAX (lane 4), DNA transfection of plasmids expressing Cas9 and EGFP-targeting sgRNA (lane 5), or Cas9 protein + EGFP-targeting sgRNA + RNAiMAX (lane 6). ( c ) T7EI assay of simultaneous genome modification at EGFP and three endogenous genes in U2OS cells 48 hours after a single treatment of 100 nM Cas9 protein, 25 nM of each of the four sgRNAs shown (100 nM total sgRNA), and 0.8 µL RNAiMAX. ( d ) Delivery of Cas9 D10A nickase and pairs of sgRNAs either by plasmid transfection or by RNAiMAX-mediated protein:sgRNA complex delivery under conditions described in ( a ) with 50 nM EGFP -disrupting sgRNAs (25 nM each) for protein delivery, and 250 ng sgRNA-expressing plasmids (125 ng each) for DNA delivery. EGFP -disrupting sgRNAs g1 + g5, or g3 + g7, are expected to result in gene disruption, while g5 + g7 target the same strand and are expected to be non-functional. ( e ) Delivery of dCas9-VP64 transcriptional activators that target NTF3 either by DNA transfection or RNAiMAX-mediated protein delivery. Error bars reflect s.d. from six biological replicates performed on different days.

    Article Snippet: The re-annealed DNA was incubated with 1 µl of T7 Endonuclease I (10 U/µl, NEB) at 37 °C for 15 min. 10 µL of 50 % glycerol was added to the T7 Endonuclease reaction and 12 µL was analyzed on a 5 % TBE 18-well Criterion PAGE gel (Bio-Rad) electrophoresed for 30 min at 200 V, then stained with 1x SYBR Gold (Life Technologies) for 30 min. Cas9-induced cleavage bands and the uncleaved band were visualized on an AlphaImager HP (Alpha Innotech) and quantified using ImageJ software .

    Techniques: Cell Culture, Variant Assay, Expressing, Flow Cytometry, Cytometry, Plasmid Preparation, Transfection, T7EI Assay, Modification, Functional Assay