t7 express lysy i  (New England Biolabs)


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    Name:
    T7 Express lysY Competent E coli High Efficiency
    Description:
    T7 Express lysY Competent E coli High Efficiency 6x0 2 ml
    Catalog Number:
    c3010i
    Price:
    173
    Size:
    1 2 ml
    Category:
    Competent Bacteria
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    Structured Review

    New England Biolabs t7 express lysy i
    T7 Express lysY Competent E coli High Efficiency
    T7 Express lysY Competent E coli High Efficiency 6x0 2 ml
    https://www.bioz.com/result/t7 express lysy i/product/New England Biolabs
    Average 95 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    t7 express lysy i - by Bioz Stars, 2020-08
    95/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Enhanced uptake of potassium or glycine betaine or export of cyclic-di-AMP restores osmoresistance in a high cyclic-di-AMP Lactococcus lactis mutant
    Article Snippet: .. Wild-type or 42 amino acid deleted busR and downstream busAA promoter were cloned into pTCV-lac and introduced into E . coli T7 Express LysY (New England Biolabs) with selection using kanamycin (50 μg/ml). .. For β-galactosidase activity assays, strains were grown overnight in LB broth without NaCl (10 g/L tryptone; 5 g/L Yeast extract) and then diluted 1:100 in the same fresh LB medium and grown at 30°C, aeration 220 rpm to early log phase (OD600 ~0.25) where 0, 0.1, 0.2, 0.3 or 0.4 M NaCl was added.

    Selection:

    Article Title: Enhanced uptake of potassium or glycine betaine or export of cyclic-di-AMP restores osmoresistance in a high cyclic-di-AMP Lactococcus lactis mutant
    Article Snippet: .. Wild-type or 42 amino acid deleted busR and downstream busAA promoter were cloned into pTCV-lac and introduced into E . coli T7 Express LysY (New England Biolabs) with selection using kanamycin (50 μg/ml). .. For β-galactosidase activity assays, strains were grown overnight in LB broth without NaCl (10 g/L tryptone; 5 g/L Yeast extract) and then diluted 1:100 in the same fresh LB medium and grown at 30°C, aeration 220 rpm to early log phase (OD600 ~0.25) where 0, 0.1, 0.2, 0.3 or 0.4 M NaCl was added.

    Isolation:

    Article Title: Context-dependent function of a conserved translational regulatory module
    Article Snippet: .. GST fusion proteins were isolated from T7 Express lysY competent E. coli (New England BioLabs) and purified using the following elution buffer: 1×PBS, 0.2% Tween 20, 150 mM NaCl, 0.1% 2-mercaptoethanol, 50 mM glutathione (reduced, pH 8.0). .. Twenty femtomoles 32 P end-labeled oligoribonucleotides (Dharmacon) were combined with GST-Cbr-PUF-2 or GST-Cbr-PUF-1.2 at various concentrations as described ( ).

    Construct:

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases
    Article Snippet: .. All constructs were expressed in T7 Express lysY/I q E . coli cells (NEB) using 0.5 mM IPTG for 2 hours at 30°C for induction. .. The β isoform of hLig3 was purified as described previously[ ].

    Article Title: Structure-function analysis of Methanobacterium thermoautotrophicum RNA ligase - engineering a thermostable ATP independent enzyme
    Article Snippet: .. The resulting constructs were sequenced and expressed in T7 Express lysY/I q E.coli cells (NEB) as follows. ..

    Purification:

    Article Title: Context-dependent function of a conserved translational regulatory module
    Article Snippet: .. GST fusion proteins were isolated from T7 Express lysY competent E. coli (New England BioLabs) and purified using the following elution buffer: 1×PBS, 0.2% Tween 20, 150 mM NaCl, 0.1% 2-mercaptoethanol, 50 mM glutathione (reduced, pH 8.0). .. Twenty femtomoles 32 P end-labeled oligoribonucleotides (Dharmacon) were combined with GST-Cbr-PUF-2 or GST-Cbr-PUF-1.2 at various concentrations as described ( ).

    Expressing:

    Article Title: Spy Go purification of SpyTag-proteins using pseudo-SpyCatcher to access an oligomerization toolbox
    Article Snippet: .. Bacterial protein expression pET28a-SpyTag-MBP, pET28a-SpyTag002-MBP, and pET28a-SpyTag-mClover3 were transformed into chemically competent E. coli BL21 (DE3) RIPL (Agilent Technologies). pET28a-scPvuII-SpyTag was transformed into T7 Express lysY /I q (NEB). pDEST14-SpyCatcher2.1 S49C E77A (SpyDock), pDEST14-SpyCatcher2.1 S49C E77X variants, pDEST14-SpyCatcher2.1 E77A N-term Cys, pDEST14-SpyCatcher2.1 S55C E77A and pDEST14-SpyCatcher002-coiled coil fusions were transformed into chemically-competent E. coli C41 (DE3), a kind gift from Anthony Watts (University of Oxford). pET28a-αDR5-SpyTag was transformed into E. coli BL21 (DE3) RIPL containing a gene encoding phosphogluconolactonase, to degrade 6-phosphogluconolactone, which promotes protein gluconoylation . ..

    Article Title: Methionine Sulfoxide Reductases Are Essential for Virulence of Salmonella Typhimurium
    Article Snippet: .. Plasmids, designated pET28c::msrA and pET28c::msrB , respectively, were transformed into E. coli T7 lysY Express (NEB) for expression of the recombinant His-tagged proteins. .. Expression of MsrA and MsrB was induced by addition of 0.5 mM IPTG at an OD600 ∼0,8, and cells were grown for another 5 h. Bacteria were harvested by centrifugation (3000×g, 25 min, 4°C) and resuspended in binding buffer containing 50 mM NaH2 PO4 , 300 mM NaCl and 10 mM imidazole (pH 8.0 for MsrA, pH 8.5 for MsrB).

    Transformation Assay:

    Article Title: Ultrahigh specificity in a network of computationally designed protein-interaction pairs
    Article Snippet: .. The genes were ligated into a linearized pET21d plasmid using Nco I and Xho I restriction sites, transformed into T7 Express lysY/I q E.coli cells (NEB), and five colonies of each design were sequenced. ..

    Article Title: Spy Go purification of SpyTag-proteins using pseudo-SpyCatcher to access an oligomerization toolbox
    Article Snippet: .. Bacterial protein expression pET28a-SpyTag-MBP, pET28a-SpyTag002-MBP, and pET28a-SpyTag-mClover3 were transformed into chemically competent E. coli BL21 (DE3) RIPL (Agilent Technologies). pET28a-scPvuII-SpyTag was transformed into T7 Express lysY /I q (NEB). pDEST14-SpyCatcher2.1 S49C E77A (SpyDock), pDEST14-SpyCatcher2.1 S49C E77X variants, pDEST14-SpyCatcher2.1 E77A N-term Cys, pDEST14-SpyCatcher2.1 S55C E77A and pDEST14-SpyCatcher002-coiled coil fusions were transformed into chemically-competent E. coli C41 (DE3), a kind gift from Anthony Watts (University of Oxford). pET28a-αDR5-SpyTag was transformed into E. coli BL21 (DE3) RIPL containing a gene encoding phosphogluconolactonase, to degrade 6-phosphogluconolactone, which promotes protein gluconoylation . ..

    Article Title: Methionine Sulfoxide Reductases Are Essential for Virulence of Salmonella Typhimurium
    Article Snippet: .. Plasmids, designated pET28c::msrA and pET28c::msrB , respectively, were transformed into E. coli T7 lysY Express (NEB) for expression of the recombinant His-tagged proteins. .. Expression of MsrA and MsrB was induced by addition of 0.5 mM IPTG at an OD600 ∼0,8, and cells were grown for another 5 h. Bacteria were harvested by centrifugation (3000×g, 25 min, 4°C) and resuspended in binding buffer containing 50 mM NaH2 PO4 , 300 mM NaCl and 10 mM imidazole (pH 8.0 for MsrA, pH 8.5 for MsrB).

    Recombinant:

    Article Title: Biosimilars: the process is the product. The example of recombinant streptokinase
    Article Snippet: .. Recombinant streptokinase variants were expressed in T7 Express lysY E. coli (NEB, Herts, UK) and cells were lysed using B-PER Direct with enzymes (Thermo Fisher Scientific, Loughborough, UK). .. Recombinant SUMO star-streptokinase fusion proteins were purified by Ni-affinity and cleaved with SUMOstar Protease I (Lifesensors) over 4 h (assessed by SDS PAGE) at 30°C in the recommended buffer.

    Article Title: Methionine Sulfoxide Reductases Are Essential for Virulence of Salmonella Typhimurium
    Article Snippet: .. Plasmids, designated pET28c::msrA and pET28c::msrB , respectively, were transformed into E. coli T7 lysY Express (NEB) for expression of the recombinant His-tagged proteins. .. Expression of MsrA and MsrB was induced by addition of 0.5 mM IPTG at an OD600 ∼0,8, and cells were grown for another 5 h. Bacteria were harvested by centrifugation (3000×g, 25 min, 4°C) and resuspended in binding buffer containing 50 mM NaH2 PO4 , 300 mM NaCl and 10 mM imidazole (pH 8.0 for MsrA, pH 8.5 for MsrB).

    Plasmid Preparation:

    Article Title: Ultrahigh specificity in a network of computationally designed protein-interaction pairs
    Article Snippet: .. The genes were ligated into a linearized pET21d plasmid using Nco I and Xho I restriction sites, transformed into T7 Express lysY/I q E.coli cells (NEB), and five colonies of each design were sequenced. ..

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  • 99
    New England Biolabs t7 endonuclease 1
    Disruption of CXCR4 in TZM-bl cells via lentiCRISPR/SaCas9 renders cells resistant to HIV-1 challenge. a CXCR4 gene disruption analysis in TZM-bl cells by the <t>T7E1</t> cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced TZM-bl cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced TZM-bl cells. Assays were performed as in Fig. 1 d. d Luciferase reporter assay to quantify HIV-1 infection level. Modified TZM-bl cells were infected with HIV-1 NL4-3 (MOI of 0.5 or 1) for 6 h, then washed three times with PBS and cultured in complete DMEM medium for 3 days. Cells were collected and lysed in 100 µl of lysis buffer (Promega) for luciferase assay. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P
    T7 Endonuclease 1, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 endonuclease 1/product/New England Biolabs
    Average 99 stars, based on 34 article reviews
    Price from $9.99 to $1999.99
    t7 endonuclease 1 - by Bioz Stars, 2020-08
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    99
    New England Biolabs co transforming bl21 t7 express lysy i q e coli
    Disruption of CXCR4 in TZM-bl cells via lentiCRISPR/SaCas9 renders cells resistant to HIV-1 challenge. a CXCR4 gene disruption analysis in TZM-bl cells by the <t>T7E1</t> cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced TZM-bl cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced TZM-bl cells. Assays were performed as in Fig. 1 d. d Luciferase reporter assay to quantify HIV-1 infection level. Modified TZM-bl cells were infected with HIV-1 NL4-3 (MOI of 0.5 or 1) for 6 h, then washed three times with PBS and cultured in complete DMEM medium for 3 days. Cells were collected and lysed in 100 µl of lysis buffer (Promega) for luciferase assay. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P
    Co Transforming Bl21 T7 Express Lysy I Q E Coli, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/co transforming bl21 t7 express lysy i q e coli/product/New England Biolabs
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    co transforming bl21 t7 express lysy i q e coli - by Bioz Stars, 2020-08
    99/100 stars
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    95
    New England Biolabs t7 express lysy i
    Disruption of CXCR4 in TZM-bl cells via lentiCRISPR/SaCas9 renders cells resistant to HIV-1 challenge. a CXCR4 gene disruption analysis in TZM-bl cells by the <t>T7E1</t> cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced TZM-bl cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced TZM-bl cells. Assays were performed as in Fig. 1 d. d Luciferase reporter assay to quantify HIV-1 infection level. Modified TZM-bl cells were infected with HIV-1 NL4-3 (MOI of 0.5 or 1) for 6 h, then washed three times with PBS and cultured in complete DMEM medium for 3 days. Cells were collected and lysed in 100 µl of lysis buffer (Promega) for luciferase assay. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P
    T7 Express Lysy I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 express lysy i/product/New England Biolabs
    Average 95 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    t7 express lysy i - by Bioz Stars, 2020-08
    95/100 stars
      Buy from Supplier

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    Disruption of CXCR4 in TZM-bl cells via lentiCRISPR/SaCas9 renders cells resistant to HIV-1 challenge. a CXCR4 gene disruption analysis in TZM-bl cells by the T7E1 cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced TZM-bl cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced TZM-bl cells. Assays were performed as in Fig. 1 d. d Luciferase reporter assay to quantify HIV-1 infection level. Modified TZM-bl cells were infected with HIV-1 NL4-3 (MOI of 0.5 or 1) for 6 h, then washed three times with PBS and cultured in complete DMEM medium for 3 days. Cells were collected and lysed in 100 µl of lysis buffer (Promega) for luciferase assay. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Journal: Retrovirology

    Article Title: Genome modification of CXCR4 by Staphylococcus aureus Cas9 renders cells resistance to HIV-1 infection

    doi: 10.1186/s12977-017-0375-0

    Figure Lengend Snippet: Disruption of CXCR4 in TZM-bl cells via lentiCRISPR/SaCas9 renders cells resistant to HIV-1 challenge. a CXCR4 gene disruption analysis in TZM-bl cells by the T7E1 cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced TZM-bl cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced TZM-bl cells. Assays were performed as in Fig. 1 d. d Luciferase reporter assay to quantify HIV-1 infection level. Modified TZM-bl cells were infected with HIV-1 NL4-3 (MOI of 0.5 or 1) for 6 h, then washed three times with PBS and cultured in complete DMEM medium for 3 days. Cells were collected and lysed in 100 µl of lysis buffer (Promega) for luciferase assay. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Article Snippet: The PCR products were digested with T7 endonuclease 1 (NEB) and resolved by 1.5% agarose gel electrophoresis.

    Techniques: Cleavage Assay, Flow Cytometry, Cytometry, Expressing, Luciferase, Reporter Assay, Infection, Modification, Cell Culture, Lysis

    AAV-mediated SaCas9/sgRNAs delivery suppresses HIV-1 infection in human primary CD4 + T cells. a A schematic diagram of AAV transfer vectors containing SaCas9 endonuclease and sgRNA. b T7E1 cleavage assay after 5 days post-transduction. c DNA sequences of CXCR4 of the AAV transduced CD4 + T cells. d Flow cytometry analysis of CXCR4 expression in AAV transduced CD4 + T cells. Assays were performed as in Fig. 1 c. e CD4 + T cells counts at different times after transduction. CXCR4 disrupted cells continued to grow normally. f Flow cytometry analysis of apoptosis following AAV delivery SaCas9/sgRNA. g HIV-1 p24 was detected in the supernatants of the CD4 + T cells treated with AAV delivered SaCas9/sgRNA following HIV-1 NL4-3 infection. For ( d , e and f ), one representative out of three independent experiments is shown. For ( g ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Journal: Retrovirology

    Article Title: Genome modification of CXCR4 by Staphylococcus aureus Cas9 renders cells resistance to HIV-1 infection

    doi: 10.1186/s12977-017-0375-0

    Figure Lengend Snippet: AAV-mediated SaCas9/sgRNAs delivery suppresses HIV-1 infection in human primary CD4 + T cells. a A schematic diagram of AAV transfer vectors containing SaCas9 endonuclease and sgRNA. b T7E1 cleavage assay after 5 days post-transduction. c DNA sequences of CXCR4 of the AAV transduced CD4 + T cells. d Flow cytometry analysis of CXCR4 expression in AAV transduced CD4 + T cells. Assays were performed as in Fig. 1 c. e CD4 + T cells counts at different times after transduction. CXCR4 disrupted cells continued to grow normally. f Flow cytometry analysis of apoptosis following AAV delivery SaCas9/sgRNA. g HIV-1 p24 was detected in the supernatants of the CD4 + T cells treated with AAV delivered SaCas9/sgRNA following HIV-1 NL4-3 infection. For ( d , e and f ), one representative out of three independent experiments is shown. For ( g ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Article Snippet: The PCR products were digested with T7 endonuclease 1 (NEB) and resolved by 1.5% agarose gel electrophoresis.

    Techniques: Infection, Cleavage Assay, Transduction, Flow Cytometry, Cytometry, Expressing

    Genome editing of CXCR4 in Jurkat T cells confers cell inhibition to HIV-1 infection by lentiCRISPR/SaCas9. a CXCR4 gene disruption analysis in Jurkat T cells by T7E1 cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced Jurkat T cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced Jurkat T cells. Assays were performed as in Fig. 1 d. d HIV-1 p24 was detected in the Jurkat T cells treated with SaCas9/sgRNA. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t test (*** P

    Journal: Retrovirology

    Article Title: Genome modification of CXCR4 by Staphylococcus aureus Cas9 renders cells resistance to HIV-1 infection

    doi: 10.1186/s12977-017-0375-0

    Figure Lengend Snippet: Genome editing of CXCR4 in Jurkat T cells confers cell inhibition to HIV-1 infection by lentiCRISPR/SaCas9. a CXCR4 gene disruption analysis in Jurkat T cells by T7E1 cleavage assay. Assays were performed as in Fig. 1 b. b Flow cytometry analysis of CXCR4 expression in lentivirus transduced Jurkat T cells. Assays were performed as in Fig. 1 c. c DNA sequences of CXCR4 of the transduced Jurkat T cells. Assays were performed as in Fig. 1 d. d HIV-1 p24 was detected in the Jurkat T cells treated with SaCas9/sgRNA. For ( b ), one representative out of three independent experiments is shown. For ( d ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t test (*** P

    Article Snippet: The PCR products were digested with T7 endonuclease 1 (NEB) and resolved by 1.5% agarose gel electrophoresis.

    Techniques: Inhibition, Infection, Cleavage Assay, Flow Cytometry, Cytometry, Expressing

    CXCR4 gene silencing by lentivirus mediated CRISPR/SaCas9 delivery protects GHOST-X4 cells from HIV-1 infection. a A schematic diagram of lentiviral transfer vectors containing CRISPR/SaCas9 components. LentiCRISPR v2 plasmid was modified by replacing the SpCas9 with SaCas9. Then, based on the SaCas9 PAM sequence 5′-NNGRRT-3′, we designed, synthesized and cloned the CXCR4 sgRNAs into the vector using the Bsmb1 . b CXCR4 gene disruption analysis in GHOST-X4 cells by the T7E1 cleavage assay. GHOST-X4 cells were transduced with lentiviruses (MOI of 40) in the presence of polybrene for 12 h, and the genomic DNA was extracted and used as template to amplify a CXCR4 fragment (1100 bp). Con: lentiviral vectors expressing SaCas9 only, #8 and #9: lentiviral vectors expressing SaCas9/sgRNA #8 and #9. c Flow cytometry analysis of CXCR4 expression in lentivirus transduced GHOST-X4 cells. Neg, unstained cells. Con, #8 and #9 as samples described in ( b ) stained with anti-CXCR4-PE. d DNA sequences of CXCR4 of the transduced GHOST-X4 cells. PCR products were cloned into pGEM-T Easy vector and sequenced. The PAM sequences are lined and highlighted in red; the target sequences were shown in blue; deletions are indicated with (−) and insertions with (+). N/N indicates ratio of WT or mutations to total sequenced clones. e Flow cytometry analysis of transduced GHOST-X4 cells on GFP expression 3 days post HIV-1 NL4-3 infection. Neg, no HIV-1 infection. f HIV-1 NL4-3 infection was determined by p24 level. The cultured supernatants were collected at the indicated days post infection, and HIV-1 p24 level was determined using a p24 ELISA kit. For ( c and e ), one representative out of three independent experiments is shown. For ( f ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Journal: Retrovirology

    Article Title: Genome modification of CXCR4 by Staphylococcus aureus Cas9 renders cells resistance to HIV-1 infection

    doi: 10.1186/s12977-017-0375-0

    Figure Lengend Snippet: CXCR4 gene silencing by lentivirus mediated CRISPR/SaCas9 delivery protects GHOST-X4 cells from HIV-1 infection. a A schematic diagram of lentiviral transfer vectors containing CRISPR/SaCas9 components. LentiCRISPR v2 plasmid was modified by replacing the SpCas9 with SaCas9. Then, based on the SaCas9 PAM sequence 5′-NNGRRT-3′, we designed, synthesized and cloned the CXCR4 sgRNAs into the vector using the Bsmb1 . b CXCR4 gene disruption analysis in GHOST-X4 cells by the T7E1 cleavage assay. GHOST-X4 cells were transduced with lentiviruses (MOI of 40) in the presence of polybrene for 12 h, and the genomic DNA was extracted and used as template to amplify a CXCR4 fragment (1100 bp). Con: lentiviral vectors expressing SaCas9 only, #8 and #9: lentiviral vectors expressing SaCas9/sgRNA #8 and #9. c Flow cytometry analysis of CXCR4 expression in lentivirus transduced GHOST-X4 cells. Neg, unstained cells. Con, #8 and #9 as samples described in ( b ) stained with anti-CXCR4-PE. d DNA sequences of CXCR4 of the transduced GHOST-X4 cells. PCR products were cloned into pGEM-T Easy vector and sequenced. The PAM sequences are lined and highlighted in red; the target sequences were shown in blue; deletions are indicated with (−) and insertions with (+). N/N indicates ratio of WT or mutations to total sequenced clones. e Flow cytometry analysis of transduced GHOST-X4 cells on GFP expression 3 days post HIV-1 NL4-3 infection. Neg, no HIV-1 infection. f HIV-1 NL4-3 infection was determined by p24 level. The cultured supernatants were collected at the indicated days post infection, and HIV-1 p24 level was determined using a p24 ELISA kit. For ( c and e ), one representative out of three independent experiments is shown. For ( f ), the graph represents 3 independent infection experiments and error bars represent SEM. Statistical analysis determined using unpaired t-test (*** P

    Article Snippet: The PCR products were digested with T7 endonuclease 1 (NEB) and resolved by 1.5% agarose gel electrophoresis.

    Techniques: CRISPR, Infection, Plasmid Preparation, Modification, Sequencing, Synthesized, Clone Assay, Cleavage Assay, Transduction, Expressing, Flow Cytometry, Cytometry, Staining, Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay