bglii  (Thermo Fisher)


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    Name:
    BglII 10 U µL
    Description:
    5 A ↓G A T C T 3 3 T C T A G ↑A 5 Thermo Scientific BglII restriction enzyme recognizes A GATCT sites and cuts best at 37°C in buffer O See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Note Also available as a FastDigest enzyme for rapid DNA digestion Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote For methylation sensitivity refer to product specifications
    Catalog Number:
    er0081
    Price:
    None
    Applications:
    Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher bglii
    <t>DNA</t> inversion in the UU172 element of U. parvum serotype 3 clonal variants 27815 T -K5 and M14-K16. Schematic illustration of the DNA inversion event in (A) 27815 T -K5 and (B) M14-K16. Southern blot analyses of (A′) 27815 T -K5 before (P0) and after (P3) selective pressure with Pab α-U172C and (B′) M14-K16 before and after selective pressure with Pab α-U144. Genomic DNA was digested with <t>BglII</t> and HincII and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots.
    5 A ↓G A T C T 3 3 T C T A G ↑A 5 Thermo Scientific BglII restriction enzyme recognizes A GATCT sites and cuts best at 37°C in buffer O See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Note Also available as a FastDigest enzyme for rapid DNA digestion Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote For methylation sensitivity refer to product specifications
    https://www.bioz.com/result/bglii/product/Thermo Fisher
    Average 97 stars, based on 168 article reviews
    Price from $9.99 to $1999.99
    bglii - by Bioz Stars, 2020-07
    97/100 stars

    Images

    1) Product Images from "Ureaplasma antigenic variation beyond MBA phase variation: DNA inversions generating chimeric structures and switching in expression of the MBA N-terminal paralogue UU172"

    Article Title: Ureaplasma antigenic variation beyond MBA phase variation: DNA inversions generating chimeric structures and switching in expression of the MBA N-terminal paralogue UU172

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2010.07474.x

    DNA inversion in the UU172 element of U. parvum serotype 3 clonal variants 27815 T -K5 and M14-K16. Schematic illustration of the DNA inversion event in (A) 27815 T -K5 and (B) M14-K16. Southern blot analyses of (A′) 27815 T -K5 before (P0) and after (P3) selective pressure with Pab α-U172C and (B′) M14-K16 before and after selective pressure with Pab α-U144. Genomic DNA was digested with BglII and HincII and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots.
    Figure Legend Snippet: DNA inversion in the UU172 element of U. parvum serotype 3 clonal variants 27815 T -K5 and M14-K16. Schematic illustration of the DNA inversion event in (A) 27815 T -K5 and (B) M14-K16. Southern blot analyses of (A′) 27815 T -K5 before (P0) and after (P3) selective pressure with Pab α-U172C and (B′) M14-K16 before and after selective pressure with Pab α-U144. Genomic DNA was digested with BglII and HincII and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots.

    Techniques Used: Southern Blot

    DNA inversion of the UU172 element in U. parvum serotype 3 clonal variant V892-K2. A and B. (A) Schematic illustration of the DNA inversion event and (B) Southern blot analysis of V892-K2 before (P0) and after (P3) selective pressure with Pab α-U172C. Genomic DNA was digested with BglII and EcoRV and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots. C. PCR analysis of subclones A and B with primer pairs 17/20, 17/22, 23/24 and 23/25 (see Table S3 ).
    Figure Legend Snippet: DNA inversion of the UU172 element in U. parvum serotype 3 clonal variant V892-K2. A and B. (A) Schematic illustration of the DNA inversion event and (B) Southern blot analysis of V892-K2 before (P0) and after (P3) selective pressure with Pab α-U172C. Genomic DNA was digested with BglII and EcoRV and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots. C. PCR analysis of subclones A and B with primer pairs 17/20, 17/22, 23/24 and 23/25 (see Table S3 ).

    Techniques Used: Variant Assay, Southern Blot, Polymerase Chain Reaction

    2) Product Images from "Construction and Evaluation of a Novel Internal Positive Control (IPC) for Detection of Coxiella burnetii by PCR"

    Article Title: Construction and Evaluation of a Novel Internal Positive Control (IPC) for Detection of Coxiella burnetii by PCR

    Journal: Jundishapur Journal of Microbiology

    doi: 10.5812/jjm.8849

    Schematic View of the Novel Strategy for IPC Construction Introduced in the Present Study A: TA-cloning of frag 1 (213 bp) and frag 2 (170 bp) corresponding to the initial and ending parts of the diagnostic 16SrRNA gene of C . burnetii . B: BglII digestion of pTV-frag 1 and pTV-frag 2 and creating the linearized forms. C: Ligation and cloning of both linear plasmids. As seen in the figure , the constructed plasmid No. 2 containing the IPC segment.
    Figure Legend Snippet: Schematic View of the Novel Strategy for IPC Construction Introduced in the Present Study A: TA-cloning of frag 1 (213 bp) and frag 2 (170 bp) corresponding to the initial and ending parts of the diagnostic 16SrRNA gene of C . burnetii . B: BglII digestion of pTV-frag 1 and pTV-frag 2 and creating the linearized forms. C: Ligation and cloning of both linear plasmids. As seen in the figure , the constructed plasmid No. 2 containing the IPC segment.

    Techniques Used: TA Cloning, Diagnostic Assay, Ligation, Clone Assay, Construct, Plasmid Preparation

    3) Product Images from "Novel Roles for Selected Genes in Meiotic DNA Processing"

    Article Title: Novel Roles for Selected Genes in Meiotic DNA Processing

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.0030222

    Further Characterization of VID21 , BRE1 , LGE1 , RMD11 , SGF73 , and DEF1 Mutants for these genes were made in an SK1 background. The plots on each graph represent wild type (black diamonds), rmd11Δ (white diamonds), bre1Δ (black triangles), lge1Δ (white triangles), sgf73Δ (black circles), def1Δ (white circles), and vid21Δ (black squares). Where error bars are not shown, the time courses are of individual experiments. A total of three experiments were carried out in each case and the data shown are consistent with those obtained in the other experiments. (A) The expression of IME1 , a primary transcription factor required for entry into the meiotic cell cycle was assessed. SK1 strains carrying a plasmid that expresses the lacZ reporter gene under the control of the IME1 promoter were grown for synchronous meioses and assessed for lacZ expression via β-galactosidase activity [ 92 ]. W303 MAT -a mutant strains for the above genes were assessed for G1 to S phase transition in mitosis after release from α-factor arrest [ 87 ]. (B) Pre-meiotic DNA replication was assessed for synchronized meiotic cultures by FACS and the change from 2c to 4c DNA content was plotted over time. See Figure S2 for the raw data of the FACS analysis for meiotic DNA replication. (C) DNA extractions from sporulation time courses were digested with BglII and meiotic DSB formation (DSBIII and IV) at the THR4 hotspot was assessed using Southern blotting and probing techniques [ 46 ]. See Figure S3 for the THR4 Southern blots. (D) Nuclear divisions (MI and MII) of the synchronized meiotic cultures in (A) were assessed with fluorescence microscopy using DAPI staining to visualize nuclear division. (E) DNA replication following release from α-factor arrest was assessed via FACS and the change from 1c to 2c DNA content was plotted against time. See Figure S4 for the raw data of the FACS analysis for mitotic DNA replication. (F) The budding index of cells released from α-factor synchrony was assessed by phase contrast microscopy.
    Figure Legend Snippet: Further Characterization of VID21 , BRE1 , LGE1 , RMD11 , SGF73 , and DEF1 Mutants for these genes were made in an SK1 background. The plots on each graph represent wild type (black diamonds), rmd11Δ (white diamonds), bre1Δ (black triangles), lge1Δ (white triangles), sgf73Δ (black circles), def1Δ (white circles), and vid21Δ (black squares). Where error bars are not shown, the time courses are of individual experiments. A total of three experiments were carried out in each case and the data shown are consistent with those obtained in the other experiments. (A) The expression of IME1 , a primary transcription factor required for entry into the meiotic cell cycle was assessed. SK1 strains carrying a plasmid that expresses the lacZ reporter gene under the control of the IME1 promoter were grown for synchronous meioses and assessed for lacZ expression via β-galactosidase activity [ 92 ]. W303 MAT -a mutant strains for the above genes were assessed for G1 to S phase transition in mitosis after release from α-factor arrest [ 87 ]. (B) Pre-meiotic DNA replication was assessed for synchronized meiotic cultures by FACS and the change from 2c to 4c DNA content was plotted over time. See Figure S2 for the raw data of the FACS analysis for meiotic DNA replication. (C) DNA extractions from sporulation time courses were digested with BglII and meiotic DSB formation (DSBIII and IV) at the THR4 hotspot was assessed using Southern blotting and probing techniques [ 46 ]. See Figure S3 for the THR4 Southern blots. (D) Nuclear divisions (MI and MII) of the synchronized meiotic cultures in (A) were assessed with fluorescence microscopy using DAPI staining to visualize nuclear division. (E) DNA replication following release from α-factor arrest was assessed via FACS and the change from 1c to 2c DNA content was plotted against time. See Figure S4 for the raw data of the FACS analysis for mitotic DNA replication. (F) The budding index of cells released from α-factor synchrony was assessed by phase contrast microscopy.

    Techniques Used: Expressing, Plasmid Preparation, Activity Assay, Mutagenesis, Sublimation, FACS, Southern Blot, Fluorescence, Microscopy, Staining

    4) Product Images from "Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector"

    Article Title: Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector

    Journal: Jundishapur Journal of Microbiology

    doi: 10.5812/jjm.27355

    Polymerase Chain Reaction and Restriction Enzyme Analysis A: PCR amplification of NS3/NS4A using specific primers, 1 kb DNA ladder (Vivantis, Malaysia). B: recombinant T/A cloning vector digested by BglII and SacII enzymes, showing the 2055 bp insert, 1 kb DNA ladder (Fermentas, Lithuania). C: pDisplay-NS3/NS4A plasmid digested by BglII and SacII enzymes, showing the 2055 bp insert, 1 kb DNA ladder (Fermentas, Lithuania).
    Figure Legend Snippet: Polymerase Chain Reaction and Restriction Enzyme Analysis A: PCR amplification of NS3/NS4A using specific primers, 1 kb DNA ladder (Vivantis, Malaysia). B: recombinant T/A cloning vector digested by BglII and SacII enzymes, showing the 2055 bp insert, 1 kb DNA ladder (Fermentas, Lithuania). C: pDisplay-NS3/NS4A plasmid digested by BglII and SacII enzymes, showing the 2055 bp insert, 1 kb DNA ladder (Fermentas, Lithuania).

    Techniques Used: Polymerase Chain Reaction, Amplification, Recombinant, Clone Assay, Plasmid Preparation

    5) Product Images from "Functional Diversity of Human Basic Helix-Loop-Helix Transcription Factor TCF4 Isoforms Generated by Alternative 5? Exon Usage and Splicing"

    Article Title: Functional Diversity of Human Basic Helix-Loop-Helix Transcription Factor TCF4 Isoforms Generated by Alternative 5? Exon Usage and Splicing

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0022138

    Expression of alternative TCF4 mRNAs in human tissues and brain regions. ( A ) RT-PCR analysis of TCF4 transcripts with different 5′ exons and ( B ) with alternative internal splicing. Transcripts are designated as in Figure 1C . The positions of bands respective to the transcripts encoding the full-length (FL) and Δ isoforms, − and + isoforms are indicated at the left on panel B. mRNAs with longer exon 18 (+) give rise to RT-PCR product that has a unique BglII restriction site enabling discrimination from RT-PCR products amplified from mRNAs not containing the 12 bps insert (−). House-keeping gene SDHA mRNA expression is shown at the bottom of the panel. PCR with no template was performed as a negative control (neg) with each primer pair.
    Figure Legend Snippet: Expression of alternative TCF4 mRNAs in human tissues and brain regions. ( A ) RT-PCR analysis of TCF4 transcripts with different 5′ exons and ( B ) with alternative internal splicing. Transcripts are designated as in Figure 1C . The positions of bands respective to the transcripts encoding the full-length (FL) and Δ isoforms, − and + isoforms are indicated at the left on panel B. mRNAs with longer exon 18 (+) give rise to RT-PCR product that has a unique BglII restriction site enabling discrimination from RT-PCR products amplified from mRNAs not containing the 12 bps insert (−). House-keeping gene SDHA mRNA expression is shown at the bottom of the panel. PCR with no template was performed as a negative control (neg) with each primer pair.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Negative Control

    6) Product Images from "Ureaplasma antigenic variation beyond MBA phase variation: DNA inversions generating chimeric structures and switching in expression of the MBA N-terminal paralogue UU172"

    Article Title: Ureaplasma antigenic variation beyond MBA phase variation: DNA inversions generating chimeric structures and switching in expression of the MBA N-terminal paralogue UU172

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2010.07474.x

    DNA inversion in the UU172 element of U. parvum serotype 3 clonal variants 27815 T -K5 and M14-K16. Schematic illustration of the DNA inversion event in (A) 27815 T -K5 and (B) M14-K16. Southern blot analyses of (A′) 27815 T -K5 before (P0) and after (P3) selective pressure with Pab α-U172C and (B′) M14-K16 before and after selective pressure with Pab α-U144. Genomic DNA was digested with BglII and HincII and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots.
    Figure Legend Snippet: DNA inversion in the UU172 element of U. parvum serotype 3 clonal variants 27815 T -K5 and M14-K16. Schematic illustration of the DNA inversion event in (A) 27815 T -K5 and (B) M14-K16. Southern blot analyses of (A′) 27815 T -K5 before (P0) and after (P3) selective pressure with Pab α-U172C and (B′) M14-K16 before and after selective pressure with Pab α-U144. Genomic DNA was digested with BglII and HincII and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots.

    Techniques Used: Southern Blot

    DNA inversion of the UU172 element in U. parvum serotype 3 clonal variant V892-K2. A and B. (A) Schematic illustration of the DNA inversion event and (B) Southern blot analysis of V892-K2 before (P0) and after (P3) selective pressure with Pab α-U172C. Genomic DNA was digested with BglII and EcoRV and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots. C. PCR analysis of subclones A and B with primer pairs 17/20, 17/22, 23/24 and 23/25 (see Table S3 ).
    Figure Legend Snippet: DNA inversion of the UU172 element in U. parvum serotype 3 clonal variant V892-K2. A and B. (A) Schematic illustration of the DNA inversion event and (B) Southern blot analysis of V892-K2 before (P0) and after (P3) selective pressure with Pab α-U172C. Genomic DNA was digested with BglII and EcoRV and hybridized with the DIG-11-dUTP-labelled probes (see Table S3 ) indicated below the blots. C. PCR analysis of subclones A and B with primer pairs 17/20, 17/22, 23/24 and 23/25 (see Table S3 ).

    Techniques Used: Variant Assay, Southern Blot, Polymerase Chain Reaction

    7) Product Images from "Three "hotspots" important for adenosine A2B receptor activation: a mutational analysis of transmembrane domains 4 and 5 and the second extracellular loop"

    Article Title: Three "hotspots" important for adenosine A2B receptor activation: a mutational analysis of transmembrane domains 4 and 5 and the second extracellular loop

    Journal: Purinergic Signalling

    doi: 10.1007/s11302-011-9251-x

    Snake plot of the adenosine A 2B R. Mutated residues within the A 2B R identified to result in increased constitutive activity are indicated in gray . These mutations originate from two previously described screens [ 2 , 3 ] and the TM4-EL2-TM5 screen described in the current paper. The putative disulfide bridges are indicated with dotted lines . The disulfide bridge conserved in many class A GPCRs links C78 3.25 and C171 EL2 . The non-conserved second disulfide bridge between EL1 and EL2 is based on an analogous bond in the crystal structures of the adenosine A 2A R (PDB: 3EML, 3QAK, 3YDO, and 3YDV); it links C72 EL1 and C167 EL2 . The restriction sites KpnI and BglII are indicated; they were used to obtain the fragment for random mutagenesis
    Figure Legend Snippet: Snake plot of the adenosine A 2B R. Mutated residues within the A 2B R identified to result in increased constitutive activity are indicated in gray . These mutations originate from two previously described screens [ 2 , 3 ] and the TM4-EL2-TM5 screen described in the current paper. The putative disulfide bridges are indicated with dotted lines . The disulfide bridge conserved in many class A GPCRs links C78 3.25 and C171 EL2 . The non-conserved second disulfide bridge between EL1 and EL2 is based on an analogous bond in the crystal structures of the adenosine A 2A R (PDB: 3EML, 3QAK, 3YDO, and 3YDV); it links C72 EL1 and C167 EL2 . The restriction sites KpnI and BglII are indicated; they were used to obtain the fragment for random mutagenesis

    Techniques Used: Activity Assay, Mutagenesis

    8) Product Images from "Two-dimensional intact mitochondrial DNA agarose electrophoresis reveals the structural complexity of the mammalian mitochondrial genome"

    Article Title: Two-dimensional intact mitochondrial DNA agarose electrophoresis reveals the structural complexity of the mammalian mitochondrial genome

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gks1324

    2D separation of mtDNA resolves previously undetected structural forms. (A–F) Total MEF DNA resolved by 2D-IMAGE and detected by Southern blot analysis with total mtDNA probe: ( A ) untreated; ( B ) DNA from isolated mitochondria; ( C ) topoisomerase I treated; ( D ) topoisomerase II treated; ( E ) BglII digested; and ( F ) S1 nuclease treated. Within panel (A) above 2D-IMAGE profile is a 1D lane in the correct orientation for the second dimension separation for comparison, with the following annotated species: catenanes (cat.); unit length nicked circles (1n NC); two-genome length nicked circle catenane (2n NC); CCCs; unit-length linear molecules (1n L). Arrow in panel A indicates position of CCC bar, which collapses by topoisomerase I or II treatment (black triangle in panels C and D). In panel D, the signal reduction in catenanes is completely recovered in linear molecules, all signal from CCC bar collapses onto relaxed CCC and a small amount of mtDNA at the 2n NC region that is resistant to topoisomerase II treatment is indicated by number sign. In panel E, single asterisks indicate structures potentially also identified in Figure 1 after BglII digestion, whereas open diamonds indicate additional molecules that were undetected in Figure 1 . In panel (F), single-stranded species removed by S1 nuclease treatment are indicated by black squares.
    Figure Legend Snippet: 2D separation of mtDNA resolves previously undetected structural forms. (A–F) Total MEF DNA resolved by 2D-IMAGE and detected by Southern blot analysis with total mtDNA probe: ( A ) untreated; ( B ) DNA from isolated mitochondria; ( C ) topoisomerase I treated; ( D ) topoisomerase II treated; ( E ) BglII digested; and ( F ) S1 nuclease treated. Within panel (A) above 2D-IMAGE profile is a 1D lane in the correct orientation for the second dimension separation for comparison, with the following annotated species: catenanes (cat.); unit length nicked circles (1n NC); two-genome length nicked circle catenane (2n NC); CCCs; unit-length linear molecules (1n L). Arrow in panel A indicates position of CCC bar, which collapses by topoisomerase I or II treatment (black triangle in panels C and D). In panel D, the signal reduction in catenanes is completely recovered in linear molecules, all signal from CCC bar collapses onto relaxed CCC and a small amount of mtDNA at the 2n NC region that is resistant to topoisomerase II treatment is indicated by number sign. In panel E, single asterisks indicate structures potentially also identified in Figure 1 after BglII digestion, whereas open diamonds indicate additional molecules that were undetected in Figure 1 . In panel (F), single-stranded species removed by S1 nuclease treatment are indicated by black squares.

    Techniques Used: Southern Blot, Isolation, Countercurrent Chromatography

    Separation of mtDNA into major topoisomers: catenanes, relaxed circles, linear molecules and supercoiled circles. ( A ) Treatment of total cellular DNA isolated from MEFs with BglII and topoisomerase I (Topo I). Untreated control (untr.) is shown for reference. Additional structures present after BglII digest are indicated by asterisks. ( B ) Treatment of total cellular DNA isolated from C2C12 myoblasts with topoisomerases I, II, IV and gyrase. ( C ) Treatment of total cellular DNA isolated from C2C12 myoblasts with S1 nuclease and E. coli exonuclease I. All gels were probed by Southern blot with COXI mtDNA sequence.
    Figure Legend Snippet: Separation of mtDNA into major topoisomers: catenanes, relaxed circles, linear molecules and supercoiled circles. ( A ) Treatment of total cellular DNA isolated from MEFs with BglII and topoisomerase I (Topo I). Untreated control (untr.) is shown for reference. Additional structures present after BglII digest are indicated by asterisks. ( B ) Treatment of total cellular DNA isolated from C2C12 myoblasts with topoisomerases I, II, IV and gyrase. ( C ) Treatment of total cellular DNA isolated from C2C12 myoblasts with S1 nuclease and E. coli exonuclease I. All gels were probed by Southern blot with COXI mtDNA sequence.

    Techniques Used: Isolation, Southern Blot, Sequencing

    Related Articles

    Amplification:

    Article Title: Functional Diversity of Human Basic Helix-Loop-Helix Transcription Factor TCF4 Isoforms Generated by Alternative 5? Exon Usage and Splicing
    Article Snippet: .. When indicated PCR amplified DNA was diluted three times and subjected to restriction with BglII (Fermentas). .. In situ hybridization cRNA probes were synthesized from BamHI linearized pSC-A-TCF4(10F-16R) with MAXIScript in vitro Transcription Kit and T3 polymerase (Ambion), using [α-35 S]UTP (Amersham Biosciences) for labeling.

    Plasmid Preparation:

    Article Title: Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector
    Article Snippet: .. The respective recombinant plasmid was extracted and cleaved by BglII and SacII (Fermentas, Lithuania) and inserted into the similarly digested eukaryotic expression vector pDisplay (Invitrogen, Carlsbad, CA, USA) with T4 DNA ligase (Invitrogen, Carlsbad, CA, USA) and transformed into E. coli DH5α. .. The pDisplay vector contains hemagglutinin A (HA) epitope tag in upstream and myc epitope in downstream of the cut sites which allow for the detection of the expressed recombinant proteins by immunofluorescence assay using anti-HA/myc antibodies.

    Polymerase Chain Reaction:

    Article Title: Functional Diversity of Human Basic Helix-Loop-Helix Transcription Factor TCF4 Isoforms Generated by Alternative 5? Exon Usage and Splicing
    Article Snippet: .. When indicated PCR amplified DNA was diluted three times and subjected to restriction with BglII (Fermentas). .. In situ hybridization cRNA probes were synthesized from BamHI linearized pSC-A-TCF4(10F-16R) with MAXIScript in vitro Transcription Kit and T3 polymerase (Ambion), using [α-35 S]UTP (Amersham Biosciences) for labeling.

    Molecular Weight:

    Article Title: Novel Roles for Selected Genes in Meiotic DNA Processing
    Article Snippet: .. The DNA from the indicated times after initiation of sporulation were digested with BglII then probed to detect DSBIII and DSBIV from the THR4 hotspot; mw represents the 1-kb molecular weight marker (Fermentas). (509 KB DOC) Click here for additional data file. ..

    Expressing:

    Article Title: Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector
    Article Snippet: .. The respective recombinant plasmid was extracted and cleaved by BglII and SacII (Fermentas, Lithuania) and inserted into the similarly digested eukaryotic expression vector pDisplay (Invitrogen, Carlsbad, CA, USA) with T4 DNA ligase (Invitrogen, Carlsbad, CA, USA) and transformed into E. coli DH5α. .. The pDisplay vector contains hemagglutinin A (HA) epitope tag in upstream and myc epitope in downstream of the cut sites which allow for the detection of the expressed recombinant proteins by immunofluorescence assay using anti-HA/myc antibodies.

    Marker:

    Article Title: Novel Roles for Selected Genes in Meiotic DNA Processing
    Article Snippet: .. The DNA from the indicated times after initiation of sporulation were digested with BglII then probed to detect DSBIII and DSBIV from the THR4 hotspot; mw represents the 1-kb molecular weight marker (Fermentas). (509 KB DOC) Click here for additional data file. ..

    Transformation Assay:

    Article Title: Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector
    Article Snippet: .. The respective recombinant plasmid was extracted and cleaved by BglII and SacII (Fermentas, Lithuania) and inserted into the similarly digested eukaryotic expression vector pDisplay (Invitrogen, Carlsbad, CA, USA) with T4 DNA ligase (Invitrogen, Carlsbad, CA, USA) and transformed into E. coli DH5α. .. The pDisplay vector contains hemagglutinin A (HA) epitope tag in upstream and myc epitope in downstream of the cut sites which allow for the detection of the expressed recombinant proteins by immunofluorescence assay using anti-HA/myc antibodies.

    Recombinant:

    Article Title: Transformation of the Cyanidioschyzon merolae chloroplast genome: prospects for understanding chloroplast function in extreme environments
    Article Snippet: .. The total DNA was digested with BglII restriction enzyme to form 2.5 kpb long DNA fragments of wild type ptDNA as well as 4.2 kbp long recombinant ptDNA and probed for the presence of psb A gene (Fig. ). .. The results of a semi-quantitative Southern blot exhibited both versions of plastid in the transformed lineages, pointing out heterogeneity of the obtained transformants.

    Article Title: Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector
    Article Snippet: .. The respective recombinant plasmid was extracted and cleaved by BglII and SacII (Fermentas, Lithuania) and inserted into the similarly digested eukaryotic expression vector pDisplay (Invitrogen, Carlsbad, CA, USA) with T4 DNA ligase (Invitrogen, Carlsbad, CA, USA) and transformed into E. coli DH5α. .. The pDisplay vector contains hemagglutinin A (HA) epitope tag in upstream and myc epitope in downstream of the cut sites which allow for the detection of the expressed recombinant proteins by immunofluorescence assay using anti-HA/myc antibodies.

    Derivative Assay:

    Article Title: Ureaplasma antigenic variation beyond MBA phase variation: DNA inversions generating chimeric structures and switching in expression of the MBA N-terminal paralogue UU172
    Article Snippet: .. Genomic DNA derived from DR1-K1 and M14-K16 was digested with 50 U of BglII (Fermentas Life Science) and 50 U of HincII (New England BioLabs) overnight at 37°C. .. Genomic DNA deriving from V892-K2 was digested with 50 U of BglII (Fermentas Life Sciences) and 50 U of EcoRV (Fermentas Life Sciences).

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  • 97
    Thermo Fisher bglii restriction enzyme
    A schematic representation of a double homologues recombination event between transformation vector (pCCATCH) and chloroplast genome ( a ). The sequence of the cat CH gene, under control of the C. merolae chloroplast promoter P <t>dna</t> K, (marked as white, curved rectangle ) of the dna K gene, was integrated into the chloroplast molecule at the selected position between the rpl 32 - psb A genes. The solid black rectangles represent the algal chloroplast sequence, also present in the transformation vector, flanking the cat CH cassettes and functioning as facilitators of double homologues recombination. b Verification of the insertion of the cat CH cassette into the rpl 32- psb A region of the chloroplast molecule via PCR and subsequent agarose gel electrophoresis of the PCR products. The PCR reaction was performed on the total DNA, isolated from the wild type ( WT ) and stable chloroplast transformant lines (transformed by B biolistic bombardment or P PEG method). Two sets of primers were used: 5UTF-5UTR amplifying a 4612 bp long product (marked as x) and 3UTF-3UTR amplifying a 3655 bp long product (marked as y). These two PCR products encompass the regions between cat CH gene and fragments from beyond the upstream or downstream homologous region in the transformation vector. The presence of the cat CH was confirmed by a PCR reaction with catCHStuI-catCHKpnI primers pair amplifying a 672 bp long product (marked as z). To exclude the possibility of unwarranted, random integration of the vector or its continuous presence in the cell, a fragment of the plasmid ori region was amplified with the pABB20R-pABB20L primer pair, yielding a 3420 bp long product, only in case of the vector control, thereby confirming the absence of any residual plasmid (marked as v). M molecular marker. c Southern blot hybridization of PCR products generated by the 5UTF-5UTR and 3UTF-3UTR primer pairs with the CATCH probe (being the PCR product generated with catCHStuI-catCHKpnI primers pair). The total DNA, isolated from the stable chloroplast transformants with the integrated pCCATCH, obtained via biolistic bombardment ( B ) or PEG method ( P ) was used as a templates for PCR. The correct integration of the transformation vector was confirmed by the size of apparent bands (4612 bp and 3655 bp, see the upper part ) and additionally by Southern blot hybridization with cat probe (the lower part ). d The Southern blot hybridization of <t>BglII-digested</t> total DNA with the psb A probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with BglII restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 8 µg of DNA. The DNA was transferred onto a nitrocellulose membrane and hybridized with psbA probe. The transformant-derived psbA containing band was calculated to have the length of 4.2 kbp. The WT control fragment was calculated to form a 2.5 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared psb A probe. The semiquantitative results showed two bands (pointed out with arrows ) in the P and B lines, representing the wild type ptDNA band and 1.8 kbp larger (containing P dnaK cat CH cassette) recombined ptDNA band. The presence of both types of plastids demonstrated heterogeneity of the obtained transformant lineages. e The Southern blot hybridization of EcoRI-digested total DNA with the cat CH probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with EcoRI restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 5 µg of DNA. The cat containing band was calculated to have the length of 10 kbp. Additionally, a positive control was added by digesting the transformation plasmid with EcoRI and AatII. This fragment was calculated to range ~10 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared 32 P enriched probe. The developed film shows no signal in the WT wells and one dominant signal in the transformed cells lines, confirming that the transformation vector had successfully recombined in a unique and single locus of the plastid without any unwarranted, illegitimate recombination with the genome or the plastid. The positive control is overexposed as the molar contribution of the cat sequence in the transformation vector is great deal higher than in the plastid
    Bglii Restriction Enzyme, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher bglii
    A schematic representation of a double homologues recombination event between transformation vector (pCCATCH) and chloroplast genome ( a ). The sequence of the cat CH gene, under control of the C. merolae chloroplast promoter P <t>dna</t> K, (marked as white, curved rectangle ) of the dna K gene, was integrated into the chloroplast molecule at the selected position between the rpl 32 - psb A genes. The solid black rectangles represent the algal chloroplast sequence, also present in the transformation vector, flanking the cat CH cassettes and functioning as facilitators of double homologues recombination. b Verification of the insertion of the cat CH cassette into the rpl 32- psb A region of the chloroplast molecule via PCR and subsequent agarose gel electrophoresis of the PCR products. The PCR reaction was performed on the total DNA, isolated from the wild type ( WT ) and stable chloroplast transformant lines (transformed by B biolistic bombardment or P PEG method). Two sets of primers were used: 5UTF-5UTR amplifying a 4612 bp long product (marked as x) and 3UTF-3UTR amplifying a 3655 bp long product (marked as y). These two PCR products encompass the regions between cat CH gene and fragments from beyond the upstream or downstream homologous region in the transformation vector. The presence of the cat CH was confirmed by a PCR reaction with catCHStuI-catCHKpnI primers pair amplifying a 672 bp long product (marked as z). To exclude the possibility of unwarranted, random integration of the vector or its continuous presence in the cell, a fragment of the plasmid ori region was amplified with the pABB20R-pABB20L primer pair, yielding a 3420 bp long product, only in case of the vector control, thereby confirming the absence of any residual plasmid (marked as v). M molecular marker. c Southern blot hybridization of PCR products generated by the 5UTF-5UTR and 3UTF-3UTR primer pairs with the CATCH probe (being the PCR product generated with catCHStuI-catCHKpnI primers pair). The total DNA, isolated from the stable chloroplast transformants with the integrated pCCATCH, obtained via biolistic bombardment ( B ) or PEG method ( P ) was used as a templates for PCR. The correct integration of the transformation vector was confirmed by the size of apparent bands (4612 bp and 3655 bp, see the upper part ) and additionally by Southern blot hybridization with cat probe (the lower part ). d The Southern blot hybridization of <t>BglII-digested</t> total DNA with the psb A probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with BglII restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 8 µg of DNA. The DNA was transferred onto a nitrocellulose membrane and hybridized with psbA probe. The transformant-derived psbA containing band was calculated to have the length of 4.2 kbp. The WT control fragment was calculated to form a 2.5 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared psb A probe. The semiquantitative results showed two bands (pointed out with arrows ) in the P and B lines, representing the wild type ptDNA band and 1.8 kbp larger (containing P dnaK cat CH cassette) recombined ptDNA band. The presence of both types of plastids demonstrated heterogeneity of the obtained transformant lineages. e The Southern blot hybridization of EcoRI-digested total DNA with the cat CH probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with EcoRI restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 5 µg of DNA. The cat containing band was calculated to have the length of 10 kbp. Additionally, a positive control was added by digesting the transformation plasmid with EcoRI and AatII. This fragment was calculated to range ~10 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared 32 P enriched probe. The developed film shows no signal in the WT wells and one dominant signal in the transformed cells lines, confirming that the transformation vector had successfully recombined in a unique and single locus of the plastid without any unwarranted, illegitimate recombination with the genome or the plastid. The positive control is overexposed as the molar contribution of the cat sequence in the transformation vector is great deal higher than in the plastid
    Bglii, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bglii/product/Thermo Fisher
    Average 93 stars, based on 168 article reviews
    Price from $9.99 to $1999.99
    bglii - by Bioz Stars, 2020-07
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    A schematic representation of a double homologues recombination event between transformation vector (pCCATCH) and chloroplast genome ( a ). The sequence of the cat CH gene, under control of the C. merolae chloroplast promoter P dna K, (marked as white, curved rectangle ) of the dna K gene, was integrated into the chloroplast molecule at the selected position between the rpl 32 - psb A genes. The solid black rectangles represent the algal chloroplast sequence, also present in the transformation vector, flanking the cat CH cassettes and functioning as facilitators of double homologues recombination. b Verification of the insertion of the cat CH cassette into the rpl 32- psb A region of the chloroplast molecule via PCR and subsequent agarose gel electrophoresis of the PCR products. The PCR reaction was performed on the total DNA, isolated from the wild type ( WT ) and stable chloroplast transformant lines (transformed by B biolistic bombardment or P PEG method). Two sets of primers were used: 5UTF-5UTR amplifying a 4612 bp long product (marked as x) and 3UTF-3UTR amplifying a 3655 bp long product (marked as y). These two PCR products encompass the regions between cat CH gene and fragments from beyond the upstream or downstream homologous region in the transformation vector. The presence of the cat CH was confirmed by a PCR reaction with catCHStuI-catCHKpnI primers pair amplifying a 672 bp long product (marked as z). To exclude the possibility of unwarranted, random integration of the vector or its continuous presence in the cell, a fragment of the plasmid ori region was amplified with the pABB20R-pABB20L primer pair, yielding a 3420 bp long product, only in case of the vector control, thereby confirming the absence of any residual plasmid (marked as v). M molecular marker. c Southern blot hybridization of PCR products generated by the 5UTF-5UTR and 3UTF-3UTR primer pairs with the CATCH probe (being the PCR product generated with catCHStuI-catCHKpnI primers pair). The total DNA, isolated from the stable chloroplast transformants with the integrated pCCATCH, obtained via biolistic bombardment ( B ) or PEG method ( P ) was used as a templates for PCR. The correct integration of the transformation vector was confirmed by the size of apparent bands (4612 bp and 3655 bp, see the upper part ) and additionally by Southern blot hybridization with cat probe (the lower part ). d The Southern blot hybridization of BglII-digested total DNA with the psb A probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with BglII restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 8 µg of DNA. The DNA was transferred onto a nitrocellulose membrane and hybridized with psbA probe. The transformant-derived psbA containing band was calculated to have the length of 4.2 kbp. The WT control fragment was calculated to form a 2.5 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared psb A probe. The semiquantitative results showed two bands (pointed out with arrows ) in the P and B lines, representing the wild type ptDNA band and 1.8 kbp larger (containing P dnaK cat CH cassette) recombined ptDNA band. The presence of both types of plastids demonstrated heterogeneity of the obtained transformant lineages. e The Southern blot hybridization of EcoRI-digested total DNA with the cat CH probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with EcoRI restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 5 µg of DNA. The cat containing band was calculated to have the length of 10 kbp. Additionally, a positive control was added by digesting the transformation plasmid with EcoRI and AatII. This fragment was calculated to range ~10 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared 32 P enriched probe. The developed film shows no signal in the WT wells and one dominant signal in the transformed cells lines, confirming that the transformation vector had successfully recombined in a unique and single locus of the plastid without any unwarranted, illegitimate recombination with the genome or the plastid. The positive control is overexposed as the molar contribution of the cat sequence in the transformation vector is great deal higher than in the plastid

    Journal: Plant Molecular Biology

    Article Title: Transformation of the Cyanidioschyzon merolae chloroplast genome: prospects for understanding chloroplast function in extreme environments

    doi: 10.1007/s11103-016-0554-8

    Figure Lengend Snippet: A schematic representation of a double homologues recombination event between transformation vector (pCCATCH) and chloroplast genome ( a ). The sequence of the cat CH gene, under control of the C. merolae chloroplast promoter P dna K, (marked as white, curved rectangle ) of the dna K gene, was integrated into the chloroplast molecule at the selected position between the rpl 32 - psb A genes. The solid black rectangles represent the algal chloroplast sequence, also present in the transformation vector, flanking the cat CH cassettes and functioning as facilitators of double homologues recombination. b Verification of the insertion of the cat CH cassette into the rpl 32- psb A region of the chloroplast molecule via PCR and subsequent agarose gel electrophoresis of the PCR products. The PCR reaction was performed on the total DNA, isolated from the wild type ( WT ) and stable chloroplast transformant lines (transformed by B biolistic bombardment or P PEG method). Two sets of primers were used: 5UTF-5UTR amplifying a 4612 bp long product (marked as x) and 3UTF-3UTR amplifying a 3655 bp long product (marked as y). These two PCR products encompass the regions between cat CH gene and fragments from beyond the upstream or downstream homologous region in the transformation vector. The presence of the cat CH was confirmed by a PCR reaction with catCHStuI-catCHKpnI primers pair amplifying a 672 bp long product (marked as z). To exclude the possibility of unwarranted, random integration of the vector or its continuous presence in the cell, a fragment of the plasmid ori region was amplified with the pABB20R-pABB20L primer pair, yielding a 3420 bp long product, only in case of the vector control, thereby confirming the absence of any residual plasmid (marked as v). M molecular marker. c Southern blot hybridization of PCR products generated by the 5UTF-5UTR and 3UTF-3UTR primer pairs with the CATCH probe (being the PCR product generated with catCHStuI-catCHKpnI primers pair). The total DNA, isolated from the stable chloroplast transformants with the integrated pCCATCH, obtained via biolistic bombardment ( B ) or PEG method ( P ) was used as a templates for PCR. The correct integration of the transformation vector was confirmed by the size of apparent bands (4612 bp and 3655 bp, see the upper part ) and additionally by Southern blot hybridization with cat probe (the lower part ). d The Southern blot hybridization of BglII-digested total DNA with the psb A probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with BglII restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 8 µg of DNA. The DNA was transferred onto a nitrocellulose membrane and hybridized with psbA probe. The transformant-derived psbA containing band was calculated to have the length of 4.2 kbp. The WT control fragment was calculated to form a 2.5 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared psb A probe. The semiquantitative results showed two bands (pointed out with arrows ) in the P and B lines, representing the wild type ptDNA band and 1.8 kbp larger (containing P dnaK cat CH cassette) recombined ptDNA band. The presence of both types of plastids demonstrated heterogeneity of the obtained transformant lineages. e The Southern blot hybridization of EcoRI-digested total DNA with the cat CH probe. The total DNA was isolated from both lineages ( P, B ) of the transformed cells as well as the wild type. After digesting with EcoRI restriction enzyme the total DNA was loaded onto the agarose gel in equal amounts of 5 µg of DNA. The cat containing band was calculated to have the length of 10 kbp. Additionally, a positive control was added by digesting the transformation plasmid with EcoRI and AatII. This fragment was calculated to range ~10 kbp band. The gel was transferred onto a nitrocellulose membrane and hybridized with an earlier prepared 32 P enriched probe. The developed film shows no signal in the WT wells and one dominant signal in the transformed cells lines, confirming that the transformation vector had successfully recombined in a unique and single locus of the plastid without any unwarranted, illegitimate recombination with the genome or the plastid. The positive control is overexposed as the molar contribution of the cat sequence in the transformation vector is great deal higher than in the plastid

    Article Snippet: Further, the DNA was digested with BglII restriction enzyme (Thermo, USA), cutting the plastid in the proximity of 3′ and 5′ ends of the psbA gene.

    Techniques: Transformation Assay, Plasmid Preparation, Sequencing, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Isolation, Amplification, Marker, Southern Blot, Hybridization, Generated, Derivative Assay, Positive Control