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  • 99
    New England Biolabs spei
    Over-expressing L1-ectodomain in MDA-MB-468 cells . (A) Schematic diagram of Lvv 1879 vector containing L1ED. 3350 bp L1 ectodomain fragment was amplified and inserted into Lvv 1879 via <t>SpeI</t> and <t>XhoI</t> restriction enzyme sites. The constructed lentivirus was used to infect MDA-MB-468 cells to establish a new stable cell line. (B) Immunostaining and FACS analysis of L1CAM level in MDA-MB-468-L1ED compared to mock vector infected and plain MDA-MB-468 cells. (C) TCA precipitation and western blotting examining over-expressed L1 ectodomain release in MDA-MB-468-L1ED culture medium by monoclonal antibody 5G3. The amount of cell associated L1 in pellets was probed by polyclonal antibody NCAM-L1 (C-20).
    Spei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2448 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore spei
    Minichromosome <t>DNA</t> is cut at only one site by <t>SpeI</t> or SwaI, which have seven and two cutting sites, respectively. ( A ) Circular minichromosome DNA showing SpeI, SwaI and PacI sites; SpeI fragment lengths are not shown for clarity. ( B ) Minichromosome DNA from permeabilized cells incubated with SpeI or ( C ) with SwaI, and deproteinized. Lane DNA, SwaI fragments produced in deproteinized cells. ( D ) The SwaI cleavage site mapped by gel hybridization. DNA linearized by SwaI (200 U/ml, 2 h) was isolated from a PFGE gel, cut by PacI (100 U/ml, 18 h), and the products were separated by PFGE. Lanes M, oligomers with HindIII fragments of λ DNA. The four fragments produced (∼140, 100, 72 and 29 kb) represent a mixture of the pairs produced after SwaI had cut at only one of its two sites in different minichromosomes. ( E ) The SwaI cleavage site mapped by fibre-FISH. The positions of the SwaI sites and hybridization probes (green) are shown above; the biotin-labelled probes were detected with anti-biotin antibodies (green) and BrdU-labelled DNA with anti-BrdU antibodies (red). Images show representative linear molecules from the two classes observed, which had been cut by SwaI (shown by the extremities of the molecule) at either the left (upper panel) or the right (lower panel) site on the map. The probe positions were aligned approximately considering the variable stretching during combing ( 22 , 23 ). Below, linear molecules cleaved at the single PacI site for comparison.
    Spei, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 151 article reviews
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    95
    Promega spei
    The strategy for constructing the Ad5 vector platform and shuttle vectors. A: The luciferase expression cassette with abrogated <t>SpeI</t> and <t>XbaI</t> sites (pShuttle-ΔSX-Luc) was inserted into the E1/E3 deletion region by an in vitro ligation method using
    Spei, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 613 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    spei  (Roche)
    92
    Roche spei
    Representative XbaI and <t>SpeI</t> <t>PFGE</t> profiles obtained for the 91 S. enterica serotype Typhi isolates under study. The PFGE profile numbering is indicated. The dendrograms generated by the BioNumerics software show the results of cluster analysis on the
    Spei, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher spei
    Targeted mutagenesis using the CRISPR/Cas9 system in transient expression in N. benthamiana. a Schematic representation of the structure of Niben101Scf04205Ctg025 (XT1) and Niben101Scf04551Ctg021 (XT2) (exons in grey , introns in white ) with the sequences of the target sites. Diagnostic restriction sites are underlined and the PAM sequence is shown in bold . b Comparison of the mutation efficiency of hCas9 and pcoCas9 targeting the XT2. Red arrow shows <t>SpeI</t> resistant PCR fragments only visible on the gRNA and hCas9 combination. c PCR/RE assay to detect simultaneous targeted mutations on XT1 and XT2. Red arrows show BsmBI and SpeI resistant PCR fragments amplified from N. benthamiana genomic <t>DNA.</t> d Alignment of XT1 and XT2 sequences obtained from different clones of uncleaved bands (see c ). XT1 target site appears in blue and XT2 target site in green . Red letters and dashes indicate insertions and deletions respectively
    Spei, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2413 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    New England Biolabs spei hf
    Targeted mutagenesis using the CRISPR/Cas9 system in transient expression in N. benthamiana. a Schematic representation of the structure of Niben101Scf04205Ctg025 (XT1) and Niben101Scf04551Ctg021 (XT2) (exons in grey , introns in white ) with the sequences of the target sites. Diagnostic restriction sites are underlined and the PAM sequence is shown in bold . b Comparison of the mutation efficiency of hCas9 and pcoCas9 targeting the XT2. Red arrow shows <t>SpeI</t> resistant PCR fragments only visible on the gRNA and hCas9 combination. c PCR/RE assay to detect simultaneous targeted mutations on XT1 and XT2. Red arrows show BsmBI and SpeI resistant PCR fragments amplified from N. benthamiana genomic <t>DNA.</t> d Alignment of XT1 and XT2 sequences obtained from different clones of uncleaved bands (see c ). XT1 target site appears in blue and XT2 target site in green . Red letters and dashes indicate insertions and deletions respectively
    Spei Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad spei
    Targeted mutagenesis using the CRISPR/Cas9 system in transient expression in N. benthamiana. a Schematic representation of the structure of Niben101Scf04205Ctg025 (XT1) and Niben101Scf04551Ctg021 (XT2) (exons in grey , introns in white ) with the sequences of the target sites. Diagnostic restriction sites are underlined and the PAM sequence is shown in bold . b Comparison of the mutation efficiency of hCas9 and pcoCas9 targeting the XT2. Red arrow shows <t>SpeI</t> resistant PCR fragments only visible on the gRNA and hCas9 combination. c PCR/RE assay to detect simultaneous targeted mutations on XT1 and XT2. Red arrows show BsmBI and SpeI resistant PCR fragments amplified from N. benthamiana genomic <t>DNA.</t> d Alignment of XT1 and XT2 sequences obtained from different clones of uncleaved bands (see c ). XT1 target site appears in blue and XT2 target site in green . Red letters and dashes indicate insertions and deletions respectively
    Spei, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/spei/product/Bio-Rad
    Average 93 stars, based on 102 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Over-expressing L1-ectodomain in MDA-MB-468 cells . (A) Schematic diagram of Lvv 1879 vector containing L1ED. 3350 bp L1 ectodomain fragment was amplified and inserted into Lvv 1879 via SpeI and XhoI restriction enzyme sites. The constructed lentivirus was used to infect MDA-MB-468 cells to establish a new stable cell line. (B) Immunostaining and FACS analysis of L1CAM level in MDA-MB-468-L1ED compared to mock vector infected and plain MDA-MB-468 cells. (C) TCA precipitation and western blotting examining over-expressed L1 ectodomain release in MDA-MB-468-L1ED culture medium by monoclonal antibody 5G3. The amount of cell associated L1 in pellets was probed by polyclonal antibody NCAM-L1 (C-20).

    Journal: Cancer Cell International

    Article Title: Soluble L1CAM promotes breast cancer cell adhesion and migration in vitro, but not invasion

    doi: 10.1186/1475-2867-10-34

    Figure Lengend Snippet: Over-expressing L1-ectodomain in MDA-MB-468 cells . (A) Schematic diagram of Lvv 1879 vector containing L1ED. 3350 bp L1 ectodomain fragment was amplified and inserted into Lvv 1879 via SpeI and XhoI restriction enzyme sites. The constructed lentivirus was used to infect MDA-MB-468 cells to establish a new stable cell line. (B) Immunostaining and FACS analysis of L1CAM level in MDA-MB-468-L1ED compared to mock vector infected and plain MDA-MB-468 cells. (C) TCA precipitation and western blotting examining over-expressed L1 ectodomain release in MDA-MB-468-L1ED culture medium by monoclonal antibody 5G3. The amount of cell associated L1 in pellets was probed by polyclonal antibody NCAM-L1 (C-20).

    Article Snippet: The following primers with SpeI or XhoI (NEB, Ipswich, MA) cleavage sites were used to amplify the sequence: sense, 5'-GAAACTAGTCGCCGGGAAAG-3'; antisense, 5'- GCCTCGAGGAGGGAGCC- 3'.

    Techniques: Expressing, Multiple Displacement Amplification, Plasmid Preparation, Amplification, Construct, Stable Transfection, Immunostaining, FACS, Infection, TCA Precipitation, Western Blot

    Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mobilizable Plasmids for Tunable Gene Expression in Francisella novicida

    doi: 10.3389/fcimb.2018.00284

    Figure Lengend Snippet: Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Article Snippet: The next day, colonies were grown in liquid LB supplemented with 50 μg/ml kanamycin, plasmid DNA was isolated and 250 ng of each plasmid was digested with SacI-HF and SpeI restriction enzymes (New England BioLabs) for 1 h. As control, both plasmids were additionally isolated from E. coli directly, without passaging in F. novicida , and digested identically.

    Techniques: Plasmid Preparation, Expressing, Clonogenic Cell Survival Assay, Transformation Assay, Isolation

    Minichromosome DNA is cut at only one site by SpeI or SwaI, which have seven and two cutting sites, respectively. ( A ) Circular minichromosome DNA showing SpeI, SwaI and PacI sites; SpeI fragment lengths are not shown for clarity. ( B ) Minichromosome DNA from permeabilized cells incubated with SpeI or ( C ) with SwaI, and deproteinized. Lane DNA, SwaI fragments produced in deproteinized cells. ( D ) The SwaI cleavage site mapped by gel hybridization. DNA linearized by SwaI (200 U/ml, 2 h) was isolated from a PFGE gel, cut by PacI (100 U/ml, 18 h), and the products were separated by PFGE. Lanes M, oligomers with HindIII fragments of λ DNA. The four fragments produced (∼140, 100, 72 and 29 kb) represent a mixture of the pairs produced after SwaI had cut at only one of its two sites in different minichromosomes. ( E ) The SwaI cleavage site mapped by fibre-FISH. The positions of the SwaI sites and hybridization probes (green) are shown above; the biotin-labelled probes were detected with anti-biotin antibodies (green) and BrdU-labelled DNA with anti-BrdU antibodies (red). Images show representative linear molecules from the two classes observed, which had been cut by SwaI (shown by the extremities of the molecule) at either the left (upper panel) or the right (lower panel) site on the map. The probe positions were aligned approximately considering the variable stretching during combing ( 22 , 23 ). Below, linear molecules cleaved at the single PacI site for comparison.

    Journal: Nucleic Acids Research

    Article Title: DNA of a circular minichromosome linearized by restriction enzymes or other reagents is resistant to further cleavage: an influence of chromatin topology on the accessibility of DNA

    doi: 10.1093/nar/gks723

    Figure Lengend Snippet: Minichromosome DNA is cut at only one site by SpeI or SwaI, which have seven and two cutting sites, respectively. ( A ) Circular minichromosome DNA showing SpeI, SwaI and PacI sites; SpeI fragment lengths are not shown for clarity. ( B ) Minichromosome DNA from permeabilized cells incubated with SpeI or ( C ) with SwaI, and deproteinized. Lane DNA, SwaI fragments produced in deproteinized cells. ( D ) The SwaI cleavage site mapped by gel hybridization. DNA linearized by SwaI (200 U/ml, 2 h) was isolated from a PFGE gel, cut by PacI (100 U/ml, 18 h), and the products were separated by PFGE. Lanes M, oligomers with HindIII fragments of λ DNA. The four fragments produced (∼140, 100, 72 and 29 kb) represent a mixture of the pairs produced after SwaI had cut at only one of its two sites in different minichromosomes. ( E ) The SwaI cleavage site mapped by fibre-FISH. The positions of the SwaI sites and hybridization probes (green) are shown above; the biotin-labelled probes were detected with anti-biotin antibodies (green) and BrdU-labelled DNA with anti-BrdU antibodies (red). Images show representative linear molecules from the two classes observed, which had been cut by SwaI (shown by the extremities of the molecule) at either the left (upper panel) or the right (lower panel) site on the map. The probe positions were aligned approximately considering the variable stretching during combing ( 22 , 23 ). Below, linear molecules cleaved at the single PacI site for comparison.

    Article Snippet: Restriction fragments of this DNA cut with SpeI or SwaI (100 U/ml) were separated by PFGE in 1% LMP agarose at 190 V/cm for 7 or 20 h and switch time ramped linearly from 0.4 to 6 or 0.3 to 3 s, respectively, excised from gels and purified on Ultrafree columns (Millipore).

    Techniques: Incubation, Produced, Hybridization, Isolation, Fluorescence In Situ Hybridization

    Sites of breakage of minichromosome DNA in γ-irradiated cells. ( A ) SpeI and SwaI EBV DNA fragments used as probes for PFGE gels. ( B ) Hybridization of these probes to gels of DNA from control cells ( C ) or cells irradiated with 100 Gy (Irrad) restricted by the same enzyme; the PFGE conditions differed according to the length of fragments to be detected. Arrows show fragments predicted from the minichromosome DNA sequence; the origin of the weakly-hybridizing fragments is discussed in the text. (C) Fibre-FISH; the hybridization probes and procedure were as described in Figure 2 E. Images show representative linear molecules from irradiated cells (100 Gy); probes are green and DNA is red, and the extremities of the molecule represent the site of breakage. ( D ) Distribution of the breakage site expressed as the % of 55 circular DNA molecules in which the break occurred in one of four quadrants.

    Journal: Nucleic Acids Research

    Article Title: DNA of a circular minichromosome linearized by restriction enzymes or other reagents is resistant to further cleavage: an influence of chromatin topology on the accessibility of DNA

    doi: 10.1093/nar/gks723

    Figure Lengend Snippet: Sites of breakage of minichromosome DNA in γ-irradiated cells. ( A ) SpeI and SwaI EBV DNA fragments used as probes for PFGE gels. ( B ) Hybridization of these probes to gels of DNA from control cells ( C ) or cells irradiated with 100 Gy (Irrad) restricted by the same enzyme; the PFGE conditions differed according to the length of fragments to be detected. Arrows show fragments predicted from the minichromosome DNA sequence; the origin of the weakly-hybridizing fragments is discussed in the text. (C) Fibre-FISH; the hybridization probes and procedure were as described in Figure 2 E. Images show representative linear molecules from irradiated cells (100 Gy); probes are green and DNA is red, and the extremities of the molecule represent the site of breakage. ( D ) Distribution of the breakage site expressed as the % of 55 circular DNA molecules in which the break occurred in one of four quadrants.

    Article Snippet: Restriction fragments of this DNA cut with SpeI or SwaI (100 U/ml) were separated by PFGE in 1% LMP agarose at 190 V/cm for 7 or 20 h and switch time ramped linearly from 0.4 to 6 or 0.3 to 3 s, respectively, excised from gels and purified on Ultrafree columns (Millipore).

    Techniques: Irradiation, Hybridization, Sequencing, Fluorescence In Situ Hybridization

    Data and flow illustrations representing the coupling of SPE and PCR sample preparation steps on the MGA device using elastomeric valves and flow control preset by channel design. ( a ) Flow control between SPE and PCR was accomplished by using differential

    Journal:

    Article Title: A fully integrated microfluidic genetic analysis system with sample-in-answer-out capability

    doi: 10.1073/pnas.0604663103

    Figure Lengend Snippet: Data and flow illustrations representing the coupling of SPE and PCR sample preparation steps on the MGA device using elastomeric valves and flow control preset by channel design. ( a ) Flow control between SPE and PCR was accomplished by using differential

    Article Snippet: The SPE and PCR domains, along with the syringe used to deliver master mix, were silanized by using Sigmacote (Sigma-Aldrich, St. Louis, MO).

    Techniques: Flow Cytometry, Polymerase Chain Reaction, Sample Prep

    The strategy for constructing the Ad5 vector platform and shuttle vectors. A: The luciferase expression cassette with abrogated SpeI and XbaI sites (pShuttle-ΔSX-Luc) was inserted into the E1/E3 deletion region by an in vitro ligation method using

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: A fiber-modified adenovirus co-expressing HSV-TK and Coli.NTR enhances antitumor activities in breast cancer cells

    doi:

    Figure Lengend Snippet: The strategy for constructing the Ad5 vector platform and shuttle vectors. A: The luciferase expression cassette with abrogated SpeI and XbaI sites (pShuttle-ΔSX-Luc) was inserted into the E1/E3 deletion region by an in vitro ligation method using

    Article Snippet: The gene fragment flanked by the SpeI and XbaI sites was PCR amplified with specific primers 5’-GCGCTGACTCTTAAGGACTAG-3’ (forward) and 5’-CGTCGCATGCTCCTCTAG-3’ (reverse) from the pAd-Luc vector, and cloned into the pGEM-T Easy vector (Promega, Wis, USA) to generate the second shuttle plasmid, pT-SX (shuttle 2).

    Techniques: Plasmid Preparation, Luciferase, Expressing, In Vitro, Ligation

    Representative XbaI and SpeI PFGE profiles obtained for the 91 S. enterica serotype Typhi isolates under study. The PFGE profile numbering is indicated. The dendrograms generated by the BioNumerics software show the results of cluster analysis on the

    Journal:

    Article Title: Clonal Expansion and Microevolution of Quinolone-Resistant Salmonella enterica Serotype Typhi in Vietnam from 1996 to 2004 ▿

    doi: 10.1128/JCM.00948-07

    Figure Lengend Snippet: Representative XbaI and SpeI PFGE profiles obtained for the 91 S. enterica serotype Typhi isolates under study. The PFGE profile numbering is indicated. The dendrograms generated by the BioNumerics software show the results of cluster analysis on the

    Article Snippet: PFGE of XbaI (Roche)- and SpeI (Roche)-digested genomic DNA was carried out with all 91 Nalr serotype Typhi isolates in this study, as described previously ( ).

    Techniques: Generated, Software

    Targeted mutagenesis using the CRISPR/Cas9 system in transient expression in N. benthamiana. a Schematic representation of the structure of Niben101Scf04205Ctg025 (XT1) and Niben101Scf04551Ctg021 (XT2) (exons in grey , introns in white ) with the sequences of the target sites. Diagnostic restriction sites are underlined and the PAM sequence is shown in bold . b Comparison of the mutation efficiency of hCas9 and pcoCas9 targeting the XT2. Red arrow shows SpeI resistant PCR fragments only visible on the gRNA and hCas9 combination. c PCR/RE assay to detect simultaneous targeted mutations on XT1 and XT2. Red arrows show BsmBI and SpeI resistant PCR fragments amplified from N. benthamiana genomic DNA. d Alignment of XT1 and XT2 sequences obtained from different clones of uncleaved bands (see c ). XT1 target site appears in blue and XT2 target site in green . Red letters and dashes indicate insertions and deletions respectively

    Journal: Plant Methods

    Article Title: A modular toolbox for gRNA–Cas9 genome engineering in plants based on the GoldenBraid standard

    doi: 10.1186/s13007-016-0101-2

    Figure Lengend Snippet: Targeted mutagenesis using the CRISPR/Cas9 system in transient expression in N. benthamiana. a Schematic representation of the structure of Niben101Scf04205Ctg025 (XT1) and Niben101Scf04551Ctg021 (XT2) (exons in grey , introns in white ) with the sequences of the target sites. Diagnostic restriction sites are underlined and the PAM sequence is shown in bold . b Comparison of the mutation efficiency of hCas9 and pcoCas9 targeting the XT2. Red arrow shows SpeI resistant PCR fragments only visible on the gRNA and hCas9 combination. c PCR/RE assay to detect simultaneous targeted mutations on XT1 and XT2. Red arrows show BsmBI and SpeI resistant PCR fragments amplified from N. benthamiana genomic DNA. d Alignment of XT1 and XT2 sequences obtained from different clones of uncleaved bands (see c ). XT1 target site appears in blue and XT2 target site in green . Red letters and dashes indicate insertions and deletions respectively

    Article Snippet: The resulting PCR products were purified with the QIAquick PCR purification kit (QIAGEN) following the manufacturer’s protocol and restriction reactions were set up with 500 ng of purified DNA and the corresponding restriction enzyme; BsmBI (Fermentas) for XT1 and SpeI (Fermentas) for XT2.

    Techniques: Mutagenesis, CRISPR, Expressing, Diagnostic Assay, Sequencing, Polymerase Chain Reaction, Amplification, Clone Assay