polymerase chain reaction pcr clean up system  (Promega)

 
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    Name:
    Wizard PCR Preps DNA Purification System
    Description:
    Purifies PCR products directly from reactions in 15 minutes
    Catalog Number:
    a7170
    Price:
    None
    Category:
    Nucleic Acid Extraction Analysis Nucleic Acid Extraction Clean up and Concentration
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    Structured Review

    Promega polymerase chain reaction pcr clean up system
    The H2A variant H2A.Z is not involved in INO80 related effects at stress genes. H2A.Z is not involved in delayed promoter <t>nucleosome</t> repopulation in arp8 Δ. ( A and B ) Nucleosome scanning assay of the CTT1 locus (−600 to +350 bp). Wild-type and htz1 Δ cells were treated with 0.4M NaCl for 10, 30 and 60 min or left untreated. NuSA are shown in (A) and quantification of NuSA of the CTT1 −1 and +1 nucleosomes in wild-type and htz1 Δ are shown in (B). Graphs in (B) are normalized to the untreated wild-type sample. ( C ) The stress induced transient recruitment of INO80 to the 5΄ end of the CTT1 locus is similar in wild-type and htz1 Δ mutant. Recruitment to the CTT1 5΄ region (+275) including the +1 nucleosome during osmotic stress (for indicated time points) was measured by ChIP of Ino80-TAP. ( D ) Expression of the CTT1 locus during osmotic stress is not influenced by absence of H2A.Z. Wild-type cells (BY4741), arp8 Δ, htz1 Δ and htz1 Δ arp8 Δ mutants were treated with 0.4M NaCl for indicated time points. CTT1 expression levels were quantified relative to VCX1 by <t>qRT-PCR.</t> Expression levels were normalized to the maximum expression levels in wild-type (10 min).
    Purifies PCR products directly from reactions in 15 minutes
    https://www.bioz.com/result/polymerase chain reaction pcr clean up system/product/Promega
    Average 98 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction pcr clean up system - by Bioz Stars, 2020-08
    98/100 stars

    Images

    1) Product Images from "INO80 represses osmostress induced gene expression by resetting promoter proximal nucleosomes"

    Article Title: INO80 represses osmostress induced gene expression by resetting promoter proximal nucleosomes

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw1292

    The H2A variant H2A.Z is not involved in INO80 related effects at stress genes. H2A.Z is not involved in delayed promoter nucleosome repopulation in arp8 Δ. ( A and B ) Nucleosome scanning assay of the CTT1 locus (−600 to +350 bp). Wild-type and htz1 Δ cells were treated with 0.4M NaCl for 10, 30 and 60 min or left untreated. NuSA are shown in (A) and quantification of NuSA of the CTT1 −1 and +1 nucleosomes in wild-type and htz1 Δ are shown in (B). Graphs in (B) are normalized to the untreated wild-type sample. ( C ) The stress induced transient recruitment of INO80 to the 5΄ end of the CTT1 locus is similar in wild-type and htz1 Δ mutant. Recruitment to the CTT1 5΄ region (+275) including the +1 nucleosome during osmotic stress (for indicated time points) was measured by ChIP of Ino80-TAP. ( D ) Expression of the CTT1 locus during osmotic stress is not influenced by absence of H2A.Z. Wild-type cells (BY4741), arp8 Δ, htz1 Δ and htz1 Δ arp8 Δ mutants were treated with 0.4M NaCl for indicated time points. CTT1 expression levels were quantified relative to VCX1 by qRT-PCR. Expression levels were normalized to the maximum expression levels in wild-type (10 min).
    Figure Legend Snippet: The H2A variant H2A.Z is not involved in INO80 related effects at stress genes. H2A.Z is not involved in delayed promoter nucleosome repopulation in arp8 Δ. ( A and B ) Nucleosome scanning assay of the CTT1 locus (−600 to +350 bp). Wild-type and htz1 Δ cells were treated with 0.4M NaCl for 10, 30 and 60 min or left untreated. NuSA are shown in (A) and quantification of NuSA of the CTT1 −1 and +1 nucleosomes in wild-type and htz1 Δ are shown in (B). Graphs in (B) are normalized to the untreated wild-type sample. ( C ) The stress induced transient recruitment of INO80 to the 5΄ end of the CTT1 locus is similar in wild-type and htz1 Δ mutant. Recruitment to the CTT1 5΄ region (+275) including the +1 nucleosome during osmotic stress (for indicated time points) was measured by ChIP of Ino80-TAP. ( D ) Expression of the CTT1 locus during osmotic stress is not influenced by absence of H2A.Z. Wild-type cells (BY4741), arp8 Δ, htz1 Δ and htz1 Δ arp8 Δ mutants were treated with 0.4M NaCl for indicated time points. CTT1 expression levels were quantified relative to VCX1 by qRT-PCR. Expression levels were normalized to the maximum expression levels in wild-type (10 min).

    Techniques Used: Variant Assay, Mutagenesis, Chromatin Immunoprecipitation, Expressing, Quantitative RT-PCR

    Related Articles

    Amplification:

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea
    Article Snippet: .. The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing. .. The second PCR assessment utilized 6 sets of primers that bound the regions shown in Table .

    Article Title: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR
    Article Snippet: .. After amplification, the resulting DNA fragments were extracted from PCR mixtures using Wizard PCR Preps DNA Purification System (Promega, Madison, USA). .. This purification procedure is required for separation of specific DNA fragments from primer dimers if they are formed during PCR.

    Agarose Gel Electrophoresis:

    Article Title: Culture-Independent Characterization of a Gene Responsible for Nitrogen Fixation in the Symbiotic Microbial Community in the Gut of the Termite Neotermes koshunensis
    Article Snippet: .. The PCR products were purified on a low-melting-point agarose gel by means of the Wizard PCR prep DNA purification system (Promega). .. The purified PCR products were digested with Hha I (Takara), and the lengths of the fluorescently labeled terminal restriction fragments of the PCR products were determined by electrophoresis by using an automatic sequencer, ALFred Express (Pharmacia), and analyzed by means of Fragment Manager software (Pharmacia).

    Ligation:

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Purification:

    Article Title: Potential Benefits of Sequential Inhibitor-Mutagen Treatments of RNA Virus Infections
    Article Snippet: .. Then, the plasmids were purified by Wizard PCR Preps DNA purification resin (Promega), and dissolved in RNase-free water. .. FMDV RNA was transcribed from the linearized plasmids by using the Riboprobe in vitro transcription system (Promega).

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea
    Article Snippet: .. The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing. .. The second PCR assessment utilized 6 sets of primers that bound the regions shown in Table .

    Article Title: KDNA Genetic Signatures Obtained by LSSP-PCR Analysis of Leishmania (Leishmania) infantum Isolated from the New and the Old World
    Article Snippet: .. LSSP-PCR Analysis For the production of kDNA signatures from the 447 bp L. infantum kDNA fragments, PCR products were purified by using the kit Wizard® PCR Preps – DNA purification System (Promega). .. After 10 times dilution in deionized water, 2 µL of PCR-purified products were used as template in a second amplification step under low stringency conditions by using the MC1 primer as driver.

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Article Title: Genetic Identification and Risk Factor Analysis of Asymptomatic Bacterial Colonization on Cardiovascular Implantable Electronic Devices
    Article Snippet: .. The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega) and then ligated into the pGEM-T Easy Vector (Promega). .. The ligation product was transformed into the E. coli strain JM109.

    Article Title: Culture-Independent Characterization of a Gene Responsible for Nitrogen Fixation in the Symbiotic Microbial Community in the Gut of the Termite Neotermes koshunensis
    Article Snippet: .. The PCR products were purified on a low-melting-point agarose gel by means of the Wizard PCR prep DNA purification system (Promega). .. The purified PCR products were digested with Hha I (Takara), and the lengths of the fluorescently labeled terminal restriction fragments of the PCR products were determined by electrophoresis by using an automatic sequencer, ALFred Express (Pharmacia), and analyzed by means of Fragment Manager software (Pharmacia).

    DNA Sequencing:

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea
    Article Snippet: .. The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing. .. The second PCR assessment utilized 6 sets of primers that bound the regions shown in Table .

    DNA Purification:

    Article Title: Potential Benefits of Sequential Inhibitor-Mutagen Treatments of RNA Virus Infections
    Article Snippet: .. Then, the plasmids were purified by Wizard PCR Preps DNA purification resin (Promega), and dissolved in RNase-free water. .. FMDV RNA was transcribed from the linearized plasmids by using the Riboprobe in vitro transcription system (Promega).

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea
    Article Snippet: .. The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing. .. The second PCR assessment utilized 6 sets of primers that bound the regions shown in Table .

    Article Title: Intramolecular Recombinations of Moloney Murine Leukemia Virus Occur during Minus-Strand DNA Synthesis
    Article Snippet: .. The remaining EDTA was removed by Wizard PCR DNA purification kit (Promega, Madison, Wis.). .. The proviruses of step 2 were amplified with primer GFP 2312 (CTGGAGTTCGTGACCG), which annealed within the 3′ end of gfp gene, and primer U3 3469 (GCTGGACCGCATCTGG), which annealed within the 3′ U3.

    Article Title: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR
    Article Snippet: .. After amplification, the resulting DNA fragments were extracted from PCR mixtures using Wizard PCR Preps DNA Purification System (Promega, Madison, USA). .. This purification procedure is required for separation of specific DNA fragments from primer dimers if they are formed during PCR.

    Article Title: KDNA Genetic Signatures Obtained by LSSP-PCR Analysis of Leishmania (Leishmania) infantum Isolated from the New and the Old World
    Article Snippet: .. LSSP-PCR Analysis For the production of kDNA signatures from the 447 bp L. infantum kDNA fragments, PCR products were purified by using the kit Wizard® PCR Preps – DNA purification System (Promega). .. After 10 times dilution in deionized water, 2 µL of PCR-purified products were used as template in a second amplification step under low stringency conditions by using the MC1 primer as driver.

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Article Title: Genetic Identification and Risk Factor Analysis of Asymptomatic Bacterial Colonization on Cardiovascular Implantable Electronic Devices
    Article Snippet: .. The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega) and then ligated into the pGEM-T Easy Vector (Promega). .. The ligation product was transformed into the E. coli strain JM109.

    Article Title: Culture-Independent Characterization of a Gene Responsible for Nitrogen Fixation in the Symbiotic Microbial Community in the Gut of the Termite Neotermes koshunensis
    Article Snippet: .. The PCR products were purified on a low-melting-point agarose gel by means of the Wizard PCR prep DNA purification system (Promega). .. The purified PCR products were digested with Hha I (Takara), and the lengths of the fluorescently labeled terminal restriction fragments of the PCR products were determined by electrophoresis by using an automatic sequencer, ALFred Express (Pharmacia), and analyzed by means of Fragment Manager software (Pharmacia).

    Polymerase Chain Reaction:

    Article Title: Potential Benefits of Sequential Inhibitor-Mutagen Treatments of RNA Virus Infections
    Article Snippet: .. Then, the plasmids were purified by Wizard PCR Preps DNA purification resin (Promega), and dissolved in RNase-free water. .. FMDV RNA was transcribed from the linearized plasmids by using the Riboprobe in vitro transcription system (Promega).

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea
    Article Snippet: .. The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing. .. The second PCR assessment utilized 6 sets of primers that bound the regions shown in Table .

    Article Title: Intramolecular Recombinations of Moloney Murine Leukemia Virus Occur during Minus-Strand DNA Synthesis
    Article Snippet: .. The remaining EDTA was removed by Wizard PCR DNA purification kit (Promega, Madison, Wis.). .. The proviruses of step 2 were amplified with primer GFP 2312 (CTGGAGTTCGTGACCG), which annealed within the 3′ end of gfp gene, and primer U3 3469 (GCTGGACCGCATCTGG), which annealed within the 3′ U3.

    Article Title: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR
    Article Snippet: .. After amplification, the resulting DNA fragments were extracted from PCR mixtures using Wizard PCR Preps DNA Purification System (Promega, Madison, USA). .. This purification procedure is required for separation of specific DNA fragments from primer dimers if they are formed during PCR.

    Article Title: KDNA Genetic Signatures Obtained by LSSP-PCR Analysis of Leishmania (Leishmania) infantum Isolated from the New and the Old World
    Article Snippet: .. LSSP-PCR Analysis For the production of kDNA signatures from the 447 bp L. infantum kDNA fragments, PCR products were purified by using the kit Wizard® PCR Preps – DNA purification System (Promega). .. After 10 times dilution in deionized water, 2 µL of PCR-purified products were used as template in a second amplification step under low stringency conditions by using the MC1 primer as driver.

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Article Title: Genetic Identification and Risk Factor Analysis of Asymptomatic Bacterial Colonization on Cardiovascular Implantable Electronic Devices
    Article Snippet: .. The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega) and then ligated into the pGEM-T Easy Vector (Promega). .. The ligation product was transformed into the E. coli strain JM109.

    Article Title: Culture-Independent Characterization of a Gene Responsible for Nitrogen Fixation in the Symbiotic Microbial Community in the Gut of the Termite Neotermes koshunensis
    Article Snippet: .. The PCR products were purified on a low-melting-point agarose gel by means of the Wizard PCR prep DNA purification system (Promega). .. The purified PCR products were digested with Hha I (Takara), and the lengths of the fluorescently labeled terminal restriction fragments of the PCR products were determined by electrophoresis by using an automatic sequencer, ALFred Express (Pharmacia), and analyzed by means of Fragment Manager software (Pharmacia).

    Transformation Assay:

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Recombinant:

    Article Title: Newcastle Disease Virus Hemagglutinin Neuraminidase as a Potential Cancer Targeting Agent
    Article Snippet: .. The ligation mixture was purified using the Wizard® PCR Preps DNA Purification Systems (Promega, USA) before the recombinant plasmids were transformed into FreeStyle™ CHO-S® cells. .. Expression of HN-acmA cassette

    Plasmid Preparation:

    Article Title: Genetic Identification and Risk Factor Analysis of Asymptomatic Bacterial Colonization on Cardiovascular Implantable Electronic Devices
    Article Snippet: .. The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega) and then ligated into the pGEM-T Easy Vector (Promega). .. The ligation product was transformed into the E. coli strain JM109.

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  • 99
    Promega pcr clean up system
    Screening and generation of Mp erf13 mutants (A) The flow chart of the screen for mutants defective in oil body formation from <t>T-DNA-insertion</t> lines. (B) A maximum likelihood phylogenetic tree of proteins with one ERF/AP2 domain. The color code is shown in Fig S1A . The branch lengths are proportional to the estimated number of substitutions per site. Bootstrap probability is indicated as a percentage on each branch with at least 50% support. A more detailed tree was presented previously ( 20 ). (C) Schematic representation of the Mp ERF13 gene structure and mutations generated in this study. Gray and black boxes indicate the UTR and coding sequences, respectively. Asterisks with numbers indicate sites of designed gRNA to generate Mp erf13-1 ge (*1 and *4) and Mp erf13-2 ge (*2 and *3). (D) <t>PCR-based</t> genotyping of Tak-1, Mp erf13 GOF , Mp erf13-1 ge , and Mp erf13-2 ge . The combinations and annealing sites of primers (a to d) are shown in (C). (E) The genomic and predicted amino acid sequences of the Mp ERF13 locus in Tak-1 and Mp erf13 mutants. Light blue, dark blue, and magenta letters indicate UTR, coding, and predicted amino acid sequences, respectively. The PAM sequences for gRNAs are underlined. The caret indicates the indel site. (F) Fluorescent and bright-field images of BODIPY-stained three-week-old thalli of Tak-1, Mp erf13-1 ge , and Mp erf13-2 ge . Bars = 0.5 mm. (G) The number of oil bodies visualized with BODIPY in a unit area (2.0 mm × 2.0 mm). Bars indicate means ± SD. Statistical analyses between Tak-1 and each genotype were conducted using a two-tailed Welch’s t -test. Sample numbers were 23 thalli for Tak-1, 24 for Mp erf13-1 ge , and 26 for Mp erf13-2 ge . p -values are 5.14×10 −11 for Mp erf13-1 ge and 5.14×10 −11 for Mp erf13-2 ge .
    Pcr Clean Up System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 3060 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr clean up system/product/Promega
    Average 99 stars, based on 3060 article reviews
    Price from $9.99 to $1999.99
    pcr clean up system - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    94
    Promega pcr clean up kit
    Amplification, sequencing and analysis of exons 2, 5, 7, and 26 . Exons 2, 5, 7, and 26 were amplified by <t>PCR</t> using primers specific to these regions (a). L-100 bp ladder. PCR products were sequenced by Sangers method. All the 4 nucleotide differences were also present at genomic <t>DNA</t> level. Representative image of exon specific amplification (b). Alignment of exon specific sequencing results with human, mouse, and rat (predicted) sequence shows 2 amino acid residue changes (c).
    Pcr Clean Up Kit, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr clean up kit/product/Promega
    Average 94 stars, based on 202 article reviews
    Price from $9.99 to $1999.99
    pcr clean up kit - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    Image Search Results


    Screening and generation of Mp erf13 mutants (A) The flow chart of the screen for mutants defective in oil body formation from T-DNA-insertion lines. (B) A maximum likelihood phylogenetic tree of proteins with one ERF/AP2 domain. The color code is shown in Fig S1A . The branch lengths are proportional to the estimated number of substitutions per site. Bootstrap probability is indicated as a percentage on each branch with at least 50% support. A more detailed tree was presented previously ( 20 ). (C) Schematic representation of the Mp ERF13 gene structure and mutations generated in this study. Gray and black boxes indicate the UTR and coding sequences, respectively. Asterisks with numbers indicate sites of designed gRNA to generate Mp erf13-1 ge (*1 and *4) and Mp erf13-2 ge (*2 and *3). (D) PCR-based genotyping of Tak-1, Mp erf13 GOF , Mp erf13-1 ge , and Mp erf13-2 ge . The combinations and annealing sites of primers (a to d) are shown in (C). (E) The genomic and predicted amino acid sequences of the Mp ERF13 locus in Tak-1 and Mp erf13 mutants. Light blue, dark blue, and magenta letters indicate UTR, coding, and predicted amino acid sequences, respectively. The PAM sequences for gRNAs are underlined. The caret indicates the indel site. (F) Fluorescent and bright-field images of BODIPY-stained three-week-old thalli of Tak-1, Mp erf13-1 ge , and Mp erf13-2 ge . Bars = 0.5 mm. (G) The number of oil bodies visualized with BODIPY in a unit area (2.0 mm × 2.0 mm). Bars indicate means ± SD. Statistical analyses between Tak-1 and each genotype were conducted using a two-tailed Welch’s t -test. Sample numbers were 23 thalli for Tak-1, 24 for Mp erf13-1 ge , and 26 for Mp erf13-2 ge . p -values are 5.14×10 −11 for Mp erf13-1 ge and 5.14×10 −11 for Mp erf13-2 ge .

    Journal: bioRxiv

    Article Title: Switching secretory pathway direction for organelle acquisition in plants

    doi: 10.1101/2020.03.02.956961

    Figure Lengend Snippet: Screening and generation of Mp erf13 mutants (A) The flow chart of the screen for mutants defective in oil body formation from T-DNA-insertion lines. (B) A maximum likelihood phylogenetic tree of proteins with one ERF/AP2 domain. The color code is shown in Fig S1A . The branch lengths are proportional to the estimated number of substitutions per site. Bootstrap probability is indicated as a percentage on each branch with at least 50% support. A more detailed tree was presented previously ( 20 ). (C) Schematic representation of the Mp ERF13 gene structure and mutations generated in this study. Gray and black boxes indicate the UTR and coding sequences, respectively. Asterisks with numbers indicate sites of designed gRNA to generate Mp erf13-1 ge (*1 and *4) and Mp erf13-2 ge (*2 and *3). (D) PCR-based genotyping of Tak-1, Mp erf13 GOF , Mp erf13-1 ge , and Mp erf13-2 ge . The combinations and annealing sites of primers (a to d) are shown in (C). (E) The genomic and predicted amino acid sequences of the Mp ERF13 locus in Tak-1 and Mp erf13 mutants. Light blue, dark blue, and magenta letters indicate UTR, coding, and predicted amino acid sequences, respectively. The PAM sequences for gRNAs are underlined. The caret indicates the indel site. (F) Fluorescent and bright-field images of BODIPY-stained three-week-old thalli of Tak-1, Mp erf13-1 ge , and Mp erf13-2 ge . Bars = 0.5 mm. (G) The number of oil bodies visualized with BODIPY in a unit area (2.0 mm × 2.0 mm). Bars indicate means ± SD. Statistical analyses between Tak-1 and each genotype were conducted using a two-tailed Welch’s t -test. Sample numbers were 23 thalli for Tak-1, 24 for Mp erf13-1 ge , and 26 for Mp erf13-2 ge . p -values are 5.14×10 −11 for Mp erf13-1 ge and 5.14×10 −11 for Mp erf13-2 ge .

    Article Snippet: After agarose gel electrophoresis of the final TAIL-PCR products, DNA bands were excised and purified using the Wizard SV Gel and PCR Clean-Up System (Promega).

    Techniques: Generated, Polymerase Chain Reaction, Staining, Two Tailed Test

    Comparison of the sensitivities for bla L1 gene detection by LAMP and conventional PCR methods. Pure genomic DNA extracted from S. maltophilia- K279a was diluted tenfold (379.0 ng/μl to 0.00379 pg/μl) and the DNA assayed by LAMP (A,B) and PCR (C) . (A) Turbidity was monitored using the Loopamp real-time turbidimeter and the OD recorded at 650 nm, at 6 s intervals. (B) Visual inspection of the color change, post-LAMP assay, and in the presence of calcein/Mn 2+ complex. (C) PCR products were analyzed by 2% agarose gel electrophoresis and stained with ethidium bromide. The DNA marker is D2000 DNA Marker (Tiangen Biotech Co., Ltd.) The size is about 179 bp.

    Journal: Frontiers in Microbiology

    Article Title: Prevalence and detection of Stenotrophomonas maltophilia carrying metallo-β-lactamase blaL1 in Beijing, China

    doi: 10.3389/fmicb.2014.00692

    Figure Lengend Snippet: Comparison of the sensitivities for bla L1 gene detection by LAMP and conventional PCR methods. Pure genomic DNA extracted from S. maltophilia- K279a was diluted tenfold (379.0 ng/μl to 0.00379 pg/μl) and the DNA assayed by LAMP (A,B) and PCR (C) . (A) Turbidity was monitored using the Loopamp real-time turbidimeter and the OD recorded at 650 nm, at 6 s intervals. (B) Visual inspection of the color change, post-LAMP assay, and in the presence of calcein/Mn 2+ complex. (C) PCR products were analyzed by 2% agarose gel electrophoresis and stained with ethidium bromide. The DNA marker is D2000 DNA Marker (Tiangen Biotech Co., Ltd.) The size is about 179 bp.

    Article Snippet: The DNA was purified with the SV GEL and PCR Clean-Up System (Promega Co., USA).

    Techniques: Polymerase Chain Reaction, Lamp Assay, Agarose Gel Electrophoresis, Staining, Marker

    Amplification, sequencing and analysis of exons 2, 5, 7, and 26 . Exons 2, 5, 7, and 26 were amplified by PCR using primers specific to these regions (a). L-100 bp ladder. PCR products were sequenced by Sangers method. All the 4 nucleotide differences were also present at genomic DNA level. Representative image of exon specific amplification (b). Alignment of exon specific sequencing results with human, mouse, and rat (predicted) sequence shows 2 amino acid residue changes (c).

    Journal: BioMed Research International

    Article Title: Developmental Testicular Expression, Cloning, and Characterization of Rat HDAC6 In Silico

    doi: 10.1155/2017/5170680

    Figure Lengend Snippet: Amplification, sequencing and analysis of exons 2, 5, 7, and 26 . Exons 2, 5, 7, and 26 were amplified by PCR using primers specific to these regions (a). L-100 bp ladder. PCR products were sequenced by Sangers method. All the 4 nucleotide differences were also present at genomic DNA level. Representative image of exon specific amplification (b). Alignment of exon specific sequencing results with human, mouse, and rat (predicted) sequence shows 2 amino acid residue changes (c).

    Article Snippet: PCR amplified DNA was cleaned using PCR clean-up kit following the kit protocol (Promega, Wisconsin, USA), sequenced, and verified using nucleotide BLAST tool.

    Techniques: Amplification, Sequencing, Polymerase Chain Reaction

    Disruption of HBV X gene via deletion using DNA fragments derived from 4 g, 8 g and 12 g cosmids in 293 cells. (A) Cleavage sites of gRNAs included in the 4 g (4gRNA, red) and 8 g (8gRNA, green) fragments. The 12 g fragment includes both 4gRNA and 8 gRNA cleavage sites. The 20‐nucleotide recognition sequences of individual guide RNAs are boxed. The coding region is indicated by thick blue lines. HBV poly(a) sequences are disrupted and replaced by chicken β‐globin poly(a) sequences to elongate the half‐life of HBV mRNAs. 18 , 21 (B) Specific cleavages using 4 g, 8 g and 12 g DNA fragments. The 293 cells were transfected with the above fragments together with the target plasmid psCM103G and the nuclear DNAs were amplified using HBV‐X F and β‐globin poly(a) R primers (shown in A), 3 days post transfection. A short exposure of the photograph of the unprocessed PCR product of 0.6 kb is also shown. Control, 293 cells transfected only with psCM103G. (C) Schematic representation of possible gRNA cleavage sites in the PCR products. Cleavage sites of 4gRNA and 8 gRNA are shown in red and green, respectively

    Journal: The Journal of Gene Medicine

    Article Title: Highly multiplex guide RNA expression units of CRISPR/Cas9 were completely stable using cosmid amplification in a novel polygonal structure, et al. Highly multiplex guide RNA expression units of CRISPR/Cas9 were completely stable using cosmid amplification in a novel polygonal structure

    doi: 10.1002/jgm.3115

    Figure Lengend Snippet: Disruption of HBV X gene via deletion using DNA fragments derived from 4 g, 8 g and 12 g cosmids in 293 cells. (A) Cleavage sites of gRNAs included in the 4 g (4gRNA, red) and 8 g (8gRNA, green) fragments. The 12 g fragment includes both 4gRNA and 8 gRNA cleavage sites. The 20‐nucleotide recognition sequences of individual guide RNAs are boxed. The coding region is indicated by thick blue lines. HBV poly(a) sequences are disrupted and replaced by chicken β‐globin poly(a) sequences to elongate the half‐life of HBV mRNAs. 18 , 21 (B) Specific cleavages using 4 g, 8 g and 12 g DNA fragments. The 293 cells were transfected with the above fragments together with the target plasmid psCM103G and the nuclear DNAs were amplified using HBV‐X F and β‐globin poly(a) R primers (shown in A), 3 days post transfection. A short exposure of the photograph of the unprocessed PCR product of 0.6 kb is also shown. Control, 293 cells transfected only with psCM103G. (C) Schematic representation of possible gRNA cleavage sites in the PCR products. Cleavage sites of 4gRNA and 8 gRNA are shown in red and green, respectively

    Article Snippet: In total, 10 μg of s‐b cosmid was digested with Sal I (Figure , asterisks) and electrophoresed overnight in a 14‐cm long 0.8% Tris‐acetate‐ethylenediaminetetraacetic acid (TAE) agarose gel at 35 V. The DNA fragment was purified using the Wizard SV Gel and PCR Clean‐up kit as described above and was self‐ligated.

    Techniques: Derivative Assay, Transfection, Plasmid Preparation, Amplification, Polymerase Chain Reaction