ex taq dna polymerase  (TaKaRa)

 
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    Name:
    TaKaRa Ex Taq DNA Polymerase
    Description:
    TaKaRa Ex Taq DNA Polymerase combines the proven performance of Takara Taq polymerase with the proofreading activity of an efficient 3 to 5 exonuclease for high sensitivity high efficiency PCR reactions It can also be used for long range PCR up to 20 kb from genomic DNA templates and up to 30 kb from lambda DNA templates Ex Taq polymerase has a higher fidelity than standard Taq with a mutation rate approximately 4 5 times lower as determined by the Kunkel method Ex Taq polymerase is supplied with optimized 10X buffer with or without Mg2 and dNTPs
    Catalog Number:
    rr001c
    Price:
    None
    Size:
    3 000 Units
    Category:
    Ex Taq polymerase Ex Taq products High yield PCR PCR
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    Structured Review

    TaKaRa ex taq dna polymerase
    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with <t>Taq</t> I. Lane M: 100 bp <t>DNA</t> ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    TaKaRa Ex Taq DNA Polymerase combines the proven performance of Takara Taq polymerase with the proofreading activity of an efficient 3 to 5 exonuclease for high sensitivity high efficiency PCR reactions It can also be used for long range PCR up to 20 kb from genomic DNA templates and up to 30 kb from lambda DNA templates Ex Taq polymerase has a higher fidelity than standard Taq with a mutation rate approximately 4 5 times lower as determined by the Kunkel method Ex Taq polymerase is supplied with optimized 10X buffer with or without Mg2 and dNTPs
    https://www.bioz.com/result/ex taq dna polymerase/product/TaKaRa
    Average 99 stars, based on 1463 article reviews
    Price from $9.99 to $1999.99
    ex taq dna polymerase - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates"

    Article Title: The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates

    Journal: Journal of Veterinary Science

    doi: 10.4142/jvs.2006.7.3.277

    PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with Taq I. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.
    Figure Legend Snippet: PCR-RFLP fragments of the 23S rRNA gene in 3% agarose gel electrophoresis digestion with Taq I. Lane M: 100 bp DNA ladder; lane 1: B. hyodysenteriae B204; lane 2: B. hyodysenteriae B234; lane 3: B. hyodysenteriae B169; lane 4: B. pilosicoli P43/6/78; lane 5 to 14: B. hyodysenteriae field isolates; lane 15: B. murdochii 56-150; lane 16: B. intermedia PWS/A; lane 17: B. innocens B256.

    Techniques Used: Polymerase Chain Reaction, Agarose Gel Electrophoresis

    2) Product Images from "Viability of an Escherichia coli pgsA Null Mutant Lacking Detectable Phosphatidylglycerol and Cardiolipin"

    Article Title: Viability of an Escherichia coli pgsA Null Mutant Lacking Detectable Phosphatidylglycerol and Cardiolipin

    Journal: Journal of Bacteriology

    doi:

    PCR analysis of the pgsA alleles of E. coli pgsA mutants. PCR products amplified with Ex Taq DNA polymerase were subjected to 1.2% agarose gel electrophoresis. Lanes 1, W3110; lanes 2, S301; lanes 3, MDL12; lanes 4 to 6, independent clones of the transductants (S330). Primer pairs FPPG5-ASFPPG1 (a) and FPPG5-ASFPPG3 (b) were used. The design and sequences of the primers are described in Materials and Methods. With the former primer pair, the wild-type pgsA allele and the pgsA :: kan allele gave products of 0.71 and 1.9 kbp, respectively. With the latter primer pair, the wild-type pgsA and the pgsA :: kan alleles gave products of 0.5 and 1.7 kbp, respectively. A DNA fragment of ca. 1.5 kbp which appeared in MDL12 (panel b, lane 3) may be the product of a false annealing of the antisense primer with a site in lacZ ′ fused to pgsA ). The molecular size markers included (two left lanes of each gel) were λ- Hin dIII digest (23.1, 9.4, 6.6, 4.4, 2.3, 2.0, and 0.56 kbp) and λ- Eco T14 I digest (19.3, 7.7, 6.2, 4.3, 3.5, 2.7, 1.9, 1.5, 0.93, and 0.42 kbp).
    Figure Legend Snippet: PCR analysis of the pgsA alleles of E. coli pgsA mutants. PCR products amplified with Ex Taq DNA polymerase were subjected to 1.2% agarose gel electrophoresis. Lanes 1, W3110; lanes 2, S301; lanes 3, MDL12; lanes 4 to 6, independent clones of the transductants (S330). Primer pairs FPPG5-ASFPPG1 (a) and FPPG5-ASFPPG3 (b) were used. The design and sequences of the primers are described in Materials and Methods. With the former primer pair, the wild-type pgsA allele and the pgsA :: kan allele gave products of 0.71 and 1.9 kbp, respectively. With the latter primer pair, the wild-type pgsA and the pgsA :: kan alleles gave products of 0.5 and 1.7 kbp, respectively. A DNA fragment of ca. 1.5 kbp which appeared in MDL12 (panel b, lane 3) may be the product of a false annealing of the antisense primer with a site in lacZ ′ fused to pgsA ). The molecular size markers included (two left lanes of each gel) were λ- Hin dIII digest (23.1, 9.4, 6.6, 4.4, 2.3, 2.0, and 0.56 kbp) and λ- Eco T14 I digest (19.3, 7.7, 6.2, 4.3, 3.5, 2.7, 1.9, 1.5, 0.93, and 0.42 kbp).

    Techniques Used: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Clone Assay

    3) Product Images from "Genomic SELEX Search for Target Promoters under the Control of the PhoQP-RstBA Signal Relay Cascade ▿"

    Article Title: Genomic SELEX Search for Target Promoters under the Control of the PhoQP-RstBA Signal Relay Cascade ▿

    Journal:

    doi: 10.1128/JB.00319-07

    RstA-dependent transcription in vitro of the asr gene. (A) Truncated linear DNA template (350 bp) containing the asr promoter was prepared by PCR using pRSasr as the template, asr-EcoRI-F and asr-BamHI-R as primers, and Ex Taq DNA polymerase (Takara)
    Figure Legend Snippet: RstA-dependent transcription in vitro of the asr gene. (A) Truncated linear DNA template (350 bp) containing the asr promoter was prepared by PCR using pRSasr as the template, asr-EcoRI-F and asr-BamHI-R as primers, and Ex Taq DNA polymerase (Takara)

    Techniques Used: In Vitro, Polymerase Chain Reaction

    4) Product Images from "Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants"

    Article Title: Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants

    Journal: BioNanoScience

    doi: 10.1007/s12668-016-0253-6

    Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.
    Figure Legend Snippet: Genotyping of Arabidopsis thaliana plants from SALK_100012 line with (a) Phusion High-Fidelity DNA polymerase; (b) Ex-Taq DNA polymerase; (c) Taq DNA polymerase; (d) Emerald Amp GT PCR Master Mix. WT - DNA from wild-type Arabidopsis plant (positive control for wild-type PCR band); 1–8 - individual plants from SALK_100012 line. Molecular weight DNA standards are shown on each side. Loading order for each DNA sample: PCR reaction for wild-type allele, PCR reaction for T-DNA mutant allele.

    Techniques Used: Polymerase Chain Reaction, Positive Control, Molecular Weight, Mutagenesis

    5) Product Images from "Novel Therapeutic Approaches for Various Cancer Types Using a Modified Sleeping Beauty-Based Gene Delivery System"

    Article Title: Novel Therapeutic Approaches for Various Cancer Types Using a Modified Sleeping Beauty-Based Gene Delivery System

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086324

    Schematic representation of the Sleeping Beauty transposon including the hTERT promoter and SV40 enhancer used in our studies. To construct the transposon plasmid, the pGL3-hT-Con and pGL3-hT-TK-Con plasmids were cut with KpnI and BamHI; the ends of the luciferase cassette and the HSV-TK cassette were filled in using DNA polymerase I and blunt end ligated into the pT-MCS vector, resulting in the pT.hTp.Con and pT.hTp.HSV-tk.Con vectors, respectively. The pT.hTp.nori.Con plasmid was constructed from the pTnori plasmid through the insertion of the human telomerase promoter (hTERTp) and the SV40 enhancer. The SV40 promoter was replaced with hTERTp by digesting the pTnori plasmid with SexAI and BglII. The SV40 enhancer was inserted between the BsmI and BamHI sites of pTnori. The hTERT promoter and the SV40 enhancer region were amplified by PCR from the pGL3-hT-Con vector using Ex-Taq polymerase.
    Figure Legend Snippet: Schematic representation of the Sleeping Beauty transposon including the hTERT promoter and SV40 enhancer used in our studies. To construct the transposon plasmid, the pGL3-hT-Con and pGL3-hT-TK-Con plasmids were cut with KpnI and BamHI; the ends of the luciferase cassette and the HSV-TK cassette were filled in using DNA polymerase I and blunt end ligated into the pT-MCS vector, resulting in the pT.hTp.Con and pT.hTp.HSV-tk.Con vectors, respectively. The pT.hTp.nori.Con plasmid was constructed from the pTnori plasmid through the insertion of the human telomerase promoter (hTERTp) and the SV40 enhancer. The SV40 promoter was replaced with hTERTp by digesting the pTnori plasmid with SexAI and BglII. The SV40 enhancer was inserted between the BsmI and BamHI sites of pTnori. The hTERT promoter and the SV40 enhancer region were amplified by PCR from the pGL3-hT-Con vector using Ex-Taq polymerase.

    Techniques Used: Construct, Plasmid Preparation, Luciferase, Amplification, Polymerase Chain Reaction

    Related Articles

    Polymerase Chain Reaction:

    Article Title: RNA sequencing of the nephron transcriptome: a technical note
    Article Snippet: .. Finally, this cDNA molecule is amplified (the first-round PCR, 18–20 cycles) using the same universal primers (UP1 and UP2) and a high-performance DNA polymerase [TaKaRa Ex Taq HS DNA polymerase (Clontech Laboratories, Mountain View, CA, USA)]. .. Once the first-round PCR is complete, a quantitative real-time PCR (qRT-PCR) for a housekeeping gene (e.g., glutaraldehyde-3-dehydrogenase, beta-actin) is performed to see if the reverse transcription and amplification are successful.

    Article Title: The Protein Phosphatase AtPP2CA Negatively Regulates Abscisic Acid Signal Transduction in Arabidopsis, and Effects of abh1 on AtPP2CA mRNA 1 mRNA 1 [W]
    Article Snippet: .. Reverse transcription (first-strand cDNA synthesis kit, Amersham Biosciences) was performed on 2.5 μ g of RNA and 2 μ L were used for PCR reactions ( Ex Taq DNA polymerase; TaKaRa Mirus Bio). .. Samples were withdrawn after 20, 24, 28, and 32 cycles (splicing) or 28, 32, and 36 cycles (T-DNA disruption lines) and products were analyzed by agarose gel electrophoresis.

    Article Title: Viability of an Escherichia coli pgsA Null Mutant Lacking Detectable Phosphatidylglycerol and Cardiolipin
    Article Snippet: .. DNA from fresh colonies of the strains to be examined was used as a template for PCR amplification with Ex Taq DNA polymerase (Takara, Tokyo, Japan). ..

    Article Title: The 23S rRNA gene PCR-RFLP used for characterization of porcine intestinal spirochete isolates
    Article Snippet: .. The PCR conditions consisted of 5 µl (50 ng/µl) of DNA and 1 µl each of primer (50 pM) in a 5 µl of 10× reaction buffer with 5 µl of 25 mM MgCl2 , 5 µl of 10 mM dNTP (each 2.5 mM) and 1 µl of 5 U Ex Taq DNA polymerase (TaKaRa, Japan) to a final volume of 50 µl on a thermal cycler (PTC-100; MJ Research, USA). .. PCR was initiated after an incubation step at 94℃ for 3 min, followed by 30 cycles of 94℃ for 30 s, 55℃ for 30 s, and 72℃ for 30 s, with a final extension step at 72℃ for 5 min. PCR products that were 517 bp were excised and purified from agarose gels using a Geneclean II Kit (Qbiogene, USA).

    Article Title: Selection of efficient Taq DNA polymerase to optimize T-DNA genotyping method for rapid detection of mutant Arabidopsis thaliana plants
    Article Snippet: .. The following DNA polymerases were tested: Phusion High-Fidelity DNA polymerase (ThermoFisher Scientific, USA, cat #F530S), Ex-Taq DNA polymerase (Clontech, USA, cat #RR001A), Taq DNA polymerase (SibEnzyme, Russia, cat #E331), Emerald Amp GT PCR Master Mix (Clontech, USA, cat #RR310A). .. 10 ng of genomic DNA and 10 μM of each primer were used in all PCR reactions; all other components of PCR reactions were added according to the manufacturer’s suggested protocol for each individual polymerase.

    Article Title: Integrating Morphology, Breeding Ground and Mitochondrial COI Gene Analysis for Species Identification of Bellamya lithophaga (Gastropoda: Viviparidae) in China
    Article Snippet: .. PCR reagents Ex-Taq DNA Polymerase, PCR buffer, MgCl2 and dNTPs (TaKaRa Biotech Co., Ltd, Dalian, China);Protease K (Merck, US-A);WizardTM DNA Clean-up System (Pro-mega, USA). ..

    Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
    Article Snippet: .. When TaKaRa Ex Taq™ polymerase was used, PCR with DNA preparation methods (A), (B), (C) and (D) generated high amount of PCR products at 40 cycles and the agarose gel electrophoresis showed clear bands of the PCR products. .. However, PCR with DNA preparation method (E) did not generate enough amount of PCR products at 40 cycles and the agarose gel electrophoresis showed no band of the PCR product ( ).

    Generated:

    Article Title: Genomic SELEX Search for Target Promoters under the Control of the PhoQP-RstBA Signal Relay Cascade ▿
    Article Snippet: .. Probes were generated by PCR amplification of the asr , csgD , nikA , yqaD , ptsP , ykfG , yecP , and gntU promoter regions by using a pair of primers, 5′-fluorescein isothiocyanate (FITC)-labeled FITCT7pro primer (5′-TAATACGACTCACTATAGGG-3′) and T7-R primer (5′-GGTTTTCCCAGTCACACGACG-3′), SELEX fragment-containing plasmids (100 ng) as the template, and Ex Taq DNA polymerase (Takara). .. PCR products with FITC at their termini were purified by PAGE.

    Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
    Article Snippet: .. When TaKaRa Ex Taq™ polymerase was used, PCR with DNA preparation methods (A), (B), (C) and (D) generated high amount of PCR products at 40 cycles and the agarose gel electrophoresis showed clear bands of the PCR products. .. However, PCR with DNA preparation method (E) did not generate enough amount of PCR products at 40 cycles and the agarose gel electrophoresis showed no band of the PCR product ( ).

    Amplification:

    Article Title: RNA sequencing of the nephron transcriptome: a technical note
    Article Snippet: .. Finally, this cDNA molecule is amplified (the first-round PCR, 18–20 cycles) using the same universal primers (UP1 and UP2) and a high-performance DNA polymerase [TaKaRa Ex Taq HS DNA polymerase (Clontech Laboratories, Mountain View, CA, USA)]. .. Once the first-round PCR is complete, a quantitative real-time PCR (qRT-PCR) for a housekeeping gene (e.g., glutaraldehyde-3-dehydrogenase, beta-actin) is performed to see if the reverse transcription and amplification are successful.

    Article Title: Viability of an Escherichia coli pgsA Null Mutant Lacking Detectable Phosphatidylglycerol and Cardiolipin
    Article Snippet: .. DNA from fresh colonies of the strains to be examined was used as a template for PCR amplification with Ex Taq DNA polymerase (Takara, Tokyo, Japan). ..

    Agarose Gel Electrophoresis:

    Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
    Article Snippet: .. When TaKaRa Ex Taq™ polymerase was used, PCR with DNA preparation methods (A), (B), (C) and (D) generated high amount of PCR products at 40 cycles and the agarose gel electrophoresis showed clear bands of the PCR products. .. However, PCR with DNA preparation method (E) did not generate enough amount of PCR products at 40 cycles and the agarose gel electrophoresis showed no band of the PCR product ( ).

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  • 99
    TaKaRa taq dna polymerase
    (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic <t>DNA.</t> PCR was performed on the isolated genomic DNA using <t>taq</t> DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for
    Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq dna polymerase/product/TaKaRa
    Average 99 stars, based on 2134 article reviews
    Price from $9.99 to $1999.99
    taq dna polymerase - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    TaKaRa pcr master mix
    cDNA sequence of mRNA de novo transcribed from <t>TNFSF9</t> in HepG2 clone 9-1 (A) HepG2 clone 9-1 cells were incubated with 500 ng/ml LPS for the indicated times. cDNA was synthesized from total RNA, and TNFSF9 expression was quantified by real-time <t>PCR</t> as described in “Materials and Methods”. (B) HepG2 cells were incubated with 500 ng/ml LPS in the presence of 5-ethynyl uridine (EU) for 16 hrs. EU-labeled RNA was isolated as described in “Materials and Methods”. TNFSF9 cDNA was amplified by PCR and the PCR products were isolated and sequenced. This experiment is representative of four independent experiments.
    Pcr Master Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1165 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr master mix/product/TaKaRa
    Average 99 stars, based on 1165 article reviews
    Price from $9.99 to $1999.99
    pcr master mix - by Bioz Stars, 2020-07
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    Image Search Results


    (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Journal: Neuroscience letters

    Article Title: Involvement of GluR2 and GluR3 subunit C-termini in the trigeminal spinal subnucleus caudalis and C1-C2 neurons in trigeminal neuropathic pain

    doi: 10.1016/j.neulet.2010.12.060

    Figure Lengend Snippet: (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Article Snippet: PCR was performed on the isolated genomic DNA using taq DNA polymerase (TaKaRa Ex Taq™, Takara, Otsu, Japan) and primer sets (primer #21–23 for GluR2 delta7: #21, 5′-ACA GAG GAA GGT AGT GGA AGG GAG-3′; #22, 5′-CTT GGT TTG GTT GTT GGT CAT AGC-3′; #23, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′; primer #31–33 for GluR3 delta7: #31, 5′-CCA ATA CTC CAC AGG GGC AAT TTA TC-3′; #32, 5′-CCG TTG ACT GTT TTG AAT CTC ACA CC-3′; #33, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′).

    Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction, Isolation

    cDNA sequence of mRNA de novo transcribed from TNFSF9 in HepG2 clone 9-1 (A) HepG2 clone 9-1 cells were incubated with 500 ng/ml LPS for the indicated times. cDNA was synthesized from total RNA, and TNFSF9 expression was quantified by real-time PCR as described in “Materials and Methods”. (B) HepG2 cells were incubated with 500 ng/ml LPS in the presence of 5-ethynyl uridine (EU) for 16 hrs. EU-labeled RNA was isolated as described in “Materials and Methods”. TNFSF9 cDNA was amplified by PCR and the PCR products were isolated and sequenced. This experiment is representative of four independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: cDNA sequence of mRNA de novo transcribed from TNFSF9 in HepG2 clone 9-1 (A) HepG2 clone 9-1 cells were incubated with 500 ng/ml LPS for the indicated times. cDNA was synthesized from total RNA, and TNFSF9 expression was quantified by real-time PCR as described in “Materials and Methods”. (B) HepG2 cells were incubated with 500 ng/ml LPS in the presence of 5-ethynyl uridine (EU) for 16 hrs. EU-labeled RNA was isolated as described in “Materials and Methods”. TNFSF9 cDNA was amplified by PCR and the PCR products were isolated and sequenced. This experiment is representative of four independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Sequencing, Incubation, Synthesized, Expressing, Real-time Polymerase Chain Reaction, Labeling, Isolation, Amplification, Polymerase Chain Reaction

    DNA sequence of genomic TNFSF9 gene edited by CRISPR-Cas9 The TNFSF9 genes in genomic DNAs from wild type and mutated HepG2 cells were amplified by PCR. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the genomic TNFSF9 sequence. On the right are shown the DNA sequences around the region of the mutated triplet in the wildtype and mutant clones. This is one representative of five independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: DNA sequence of genomic TNFSF9 gene edited by CRISPR-Cas9 The TNFSF9 genes in genomic DNAs from wild type and mutated HepG2 cells were amplified by PCR. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the genomic TNFSF9 sequence. On the right are shown the DNA sequences around the region of the mutated triplet in the wildtype and mutant clones. This is one representative of five independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

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

    Heteroduplex formation by the cDNA of de novo transcripts of TNFSF9 from HepG2 clone 9-1 Total RNA and EU-labeled RNAs were isolated from clone 9-1 HepG2 cells. cDNA was synthesized and the TNFSF9 cDNA was amplified by PCR as described in “Materials and Methods”. The PCR products were annealed and incubated in the presence or absence of T7E1 endonuclease, and the products were fractionated on 1.2% agarose gels and visualized under UV. This is one representative of three independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: Heteroduplex formation by the cDNA of de novo transcripts of TNFSF9 from HepG2 clone 9-1 Total RNA and EU-labeled RNAs were isolated from clone 9-1 HepG2 cells. cDNA was synthesized and the TNFSF9 cDNA was amplified by PCR as described in “Materials and Methods”. The PCR products were annealed and incubated in the presence or absence of T7E1 endonuclease, and the products were fractionated on 1.2% agarose gels and visualized under UV. This is one representative of three independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Labeling, Isolation, Synthesized, Amplification, Polymerase Chain Reaction, Incubation

    cDNA sequence of mRNA transcribed from mutant genomic TNFSF9 TNFSF9 cDNA was synthesized and amplified by PCR from total RNA extracted from wild type and mutated HepG2 cells. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the TNFSF9 cDNA sequences. On the right are shown the cDNA sequences around the region of the mutated triplet in the wild type and mutant clones. This is one representative of four independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: cDNA sequence of mRNA transcribed from mutant genomic TNFSF9 TNFSF9 cDNA was synthesized and amplified by PCR from total RNA extracted from wild type and mutated HepG2 cells. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the TNFSF9 cDNA sequences. On the right are shown the cDNA sequences around the region of the mutated triplet in the wild type and mutant clones. This is one representative of four independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Sequencing, Mutagenesis, Synthesized, Amplification, Polymerase Chain Reaction, Isolation, Clone Assay

    Effects of TGF-β1 on cell morphology, E-cadherin expression, and changes in N -glycans in GE11 cells. GE11 cells were grown in 6-well (2 × 10 5 ) or bottom dishes (2 × 10 4 ) for 24 h and then replaced with fresh complete medium with or without TGF-β1 (5 ng/ml) for another 4 days of incubation. A, cell morphology of the indicated cells was photographed. Photographs were taken of living cells using a ×10 objective. Scale bar, 100 μm. B, E-cadherin expressed on the cell surface was stained with anti-E-cadherin primary antibody, followed by incubation with Alexa Fluor-conjugated secondary antibody. Scale bar, 25 μm. C, total expression levels of E-cadherin were analyzed using Western blotting. Cell lysates from those cells were immunoblotted with anti-E-cadherin antibody. D, equal amounts of cell lysate proteins (20 μg) were used as the enzymatic source for the examination of GnT-III activities. S, substrate; P, product. E, mRNA expression of GnT-III . Quantitative RT-PCR was performed by monitoring in real time the increase in fluorescence of the SYBR Green dye on an ABI StepOnePlus. The mean number of cycles to the threshold ( CT ) of fluorescence detection was calculated for each sample, and the results were normalized to the mean CT of GAPDH for each sample tested. The changes in N -glycans were detected by E4-PHA ( F ) and concanavalin A ( ConA ) ( G ) lectin blot. α-Tubulin was used as a load control. H, N -glycans of GE11 cells cultured under normal conditions and treated with ( bottom ) or without ( top ) TGF-β for 4 days were released with peptide: N -glycosidase F ( PNGaseF ), as described under “Experimental Procedures,” digested with sialidase, and subjected to reversed-phase HPLC. The elution times for those PA-bisected N -glycans were compared with standard PA- N -glycans.

    Journal: The Journal of Biological Chemistry

    Article Title: Roles of N-Acetylglucosaminyltransferase III in Epithelial-to-Mesenchymal Transition Induced by Transforming Growth Factor ?1 (TGF-?1) in Epithelial Cell Lines *

    doi: 10.1074/jbc.M111.262154

    Figure Lengend Snippet: Effects of TGF-β1 on cell morphology, E-cadherin expression, and changes in N -glycans in GE11 cells. GE11 cells were grown in 6-well (2 × 10 5 ) or bottom dishes (2 × 10 4 ) for 24 h and then replaced with fresh complete medium with or without TGF-β1 (5 ng/ml) for another 4 days of incubation. A, cell morphology of the indicated cells was photographed. Photographs were taken of living cells using a ×10 objective. Scale bar, 100 μm. B, E-cadherin expressed on the cell surface was stained with anti-E-cadherin primary antibody, followed by incubation with Alexa Fluor-conjugated secondary antibody. Scale bar, 25 μm. C, total expression levels of E-cadherin were analyzed using Western blotting. Cell lysates from those cells were immunoblotted with anti-E-cadherin antibody. D, equal amounts of cell lysate proteins (20 μg) were used as the enzymatic source for the examination of GnT-III activities. S, substrate; P, product. E, mRNA expression of GnT-III . Quantitative RT-PCR was performed by monitoring in real time the increase in fluorescence of the SYBR Green dye on an ABI StepOnePlus. The mean number of cycles to the threshold ( CT ) of fluorescence detection was calculated for each sample, and the results were normalized to the mean CT of GAPDH for each sample tested. The changes in N -glycans were detected by E4-PHA ( F ) and concanavalin A ( ConA ) ( G ) lectin blot. α-Tubulin was used as a load control. H, N -glycans of GE11 cells cultured under normal conditions and treated with ( bottom ) or without ( top ) TGF-β for 4 days were released with peptide: N -glycosidase F ( PNGaseF ), as described under “Experimental Procedures,” digested with sialidase, and subjected to reversed-phase HPLC. The elution times for those PA-bisected N -glycans were compared with standard PA- N -glycans.

    Article Snippet: Real time PCR was performed with a StepOnePlus real time PCR system (Applied Biosystems, Inc., Foster City, CA) using SYBR® Premix Ex TaqTM II PCR master mixes (Takara, Japan).

    Techniques: Expressing, Incubation, Staining, Western Blot, Quantitative RT-PCR, Fluorescence, SYBR Green Assay, Cell Culture, High Performance Liquid Chromatography

    Expression levels of miR-192, -194 and -215 in three colorectal cancer (CRC) cell lines (HT-29, HCT-116 and SW-620). Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Data are presented in CRC cell lines relative to normal colorectal tissues

    Journal: Experimental and Therapeutic Medicine

    Article Title: microRNA-192, -194 and -215 are frequently downregulated in colorectal cancer

    doi: 10.3892/etm.2011.436

    Figure Lengend Snippet: Expression levels of miR-192, -194 and -215 in three colorectal cancer (CRC) cell lines (HT-29, HCT-116 and SW-620). Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Data are presented in CRC cell lines relative to normal colorectal tissues

    Article Snippet: According to the manufacturer's instructions, real-time PCR was performed using the SYBR Premix Ex Taq™ II kit (Takara Bio, Kyoto, Japan) with a Rotor-gene 6000 system (Qiagen, Valencia, CA, USA) ( ).

    Techniques: Expressing

    Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented

    Journal: Experimental and Therapeutic Medicine

    Article Title: microRNA-192, -194 and -215 are frequently downregulated in colorectal cancer

    doi: 10.3892/etm.2011.436

    Figure Lengend Snippet: Expression levels of miR-192, -194 and -215 in 107 patients with colorectal cancer (CRC). (A, C and E) Quantification of miRNAs was measured by SYBR Premix Ex Taq II. Each sample was analyzed in triplicate and repeated three times. Data are presented

    Article Snippet: According to the manufacturer's instructions, real-time PCR was performed using the SYBR Premix Ex Taq™ II kit (Takara Bio, Kyoto, Japan) with a Rotor-gene 6000 system (Qiagen, Valencia, CA, USA) ( ).

    Techniques: Expressing