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Stratagene herculase dnapol
Herculase Dnapol, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/herculase dnapol/product/Stratagene
Average 85 stars, based on 2 article reviews
Price from $9.99 to $1999.99
herculase dnapol - by Bioz Stars, 2020-04
85/100 stars

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Clone Assay:

Article Title: STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites
Article Snippet: PCR was performed using Herculase DNA polymerase (Stratagene). .. PCR primers were obtained from Integrated DNA Technologies and their sequence is listed in . mCh-STIM1 (kind gift from R. Lewis, Stanford, CA) and all truncated and internal deletion were cloned into the EcoR I and Xho I restriction sites of pCDNA3.1+.

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: Plasmid DNA from clones containing inserts of the expected size were scaled up using Qiagen maxiprep kits. .. The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene).

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: Paragraph title: Cloning and mutagenesis ... UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene.

Amplification:

Article Title: H2A.B facilitates transcription elongation at methylated CpG loci
Article Snippet: Genomic DNA associated with H2A.B was collected by ChIP and amplified by PCR. .. For region C of Kcnq1 , Herculase DNA polymerase (Stratagene) was used for the PCR, with 1× Herculase buffer, 0.3 μM of each primer, 0.8 mM deoxynucleoside triphosphates, 4% dimethyl sulfoxide, and 0.1 μCi of [32 P]dCTP.

Article Title: Identification of a Porphyromonas gingivalis Receptor for the Streptococcus gordonii SspB Protein
Article Snippet: .. Annealing of the primer and RNA was carried out at 72°C for 2 min and then at 48°C for 1 h. The resulting cDNA was amplified, with each 100 μl of PCR mixture containing 1× PCR buffer, 3 μl of cDNA, 1.5 mM MgCl2 , 10 mM deoxynucleoside triphosphate, 100 ng of each primer, and 2.5 U of Herculase DNA polymerase (Stratagene). .. To identify molecules of P. gingivalis that interact with SspB protein, surface proteins were first labeled with biotin and extracted with EDTA and mild sonication.

Article Title: hERG1 channels are overexpressed in glioblastoma multiforme and modulate VEGF secretion in glioblastoma cell lines
Article Snippet: .. Total cDNA was used as a template for PCR amplification with Herculase DNApol (Stratagene, Cedar Creek, TX) and each of the primer pairs reported below. .. As a quality control on cDNA, PCR amplification of the messenger relative to the housekeeping gene gapdh was performed using the following primer pair: Fw: 5′-AACAGCCTCAAGATCATCAGCAA-3′ Rev: 5′-CAGTCTGGGTGGCAGTGAT-3′ (NG 003027, nucleotides 457–564) Samples positive to gapdh amplification were further analysed using the following primers: hERG1 fw: 5′-TCCAGCGGCTGTACTCGGGC-3′ hERG1 rev: 5′-TGGACCAGAAGTGGTCGGAGAACTC-3′ (U04270, nucleotides 2171–2746). helk2 fw: 5′-CTGCCCTGCGGGGCACTGTCTG-3′ helk2 rev: 5′-AGATCTGGGGGCACATTCCATG-3′ (NM012284 nucleotides 1802–2516).

Construct:

Article Title: STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites
Article Snippet: PCR was performed using Herculase DNA polymerase (Stratagene). .. R. Lewis also kindly provided the HRP-STIM1 construct. pGFP-Sec61ß and pGFP-Rtn3C were obtained from T. Rapoport (Harvard Medical School, MA) and Orai1-GFP was obtained from B. Baird (Cornell University, NY).

Incubation:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene). .. To form holoenzyme to be used in the footprinting experiments, core RNA polymerase was incubated with a threefold molar excess of the FeBABE-derivatized σA on ice for 30 min.

Expressing:

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene. .. PCR products were cloned into pCR4-TOPO (Invitrogen) and subsequently subcloned into the NcoI and XhoI sites of p415-GPD (a low-copy yeast expression vector with a constitutive GPD promoter; AmpR , LEU2 marker, and CEN/ARS origin of replication) ( ).

Transformation Assay:

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: The PCR product was cloned into the digested vector and the resulting plasmid was transformed into E. coli XL1 blue cells (Stratagene). .. The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene).

Conjugation Assay:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: The efficiency of conjugation was determined by estimating free side chains of both conjugated and unconjugated proteins with the fluorescent reagent CPM [7-diethylamino-3-(4′maleimidylphenyl)-4-methylcoumarin] (Molecular Probes) as described previously ( ). .. The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene).

Ligation:

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). .. The PCR product was agarose gel purified using a Qiaquick gel extraction kit, and a series of ligation reactions using increasing amounts of the eluted PCR product were set up overnight at 14 °C.

Footprinting:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: Paragraph title: Protein preparation and in vitro footprinting assays. ... The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene).

Article Title: Spo0A-Dependent Activation of an Extended -10 Region Promoter in Bacillus subtilis
Article Snippet: Paragraph title: Preparation of end-labeled DNA and DNase I footprinting. ... The purified labeled probe was used in a PCR containing 36 pmol of unlabeled SKF4Rev primer and Herculase DNA polymerase (Stratagene).

Introduce:

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: Synthetic oligonucleotide primers for mutagenic PCR were designed to introduce the AKAS insert of the Scott lab’s f1.K into the M13KO7 major coat protein (P8). .. A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Identification of a Porphyromonas gingivalis Receptor for the Streptococcus gordonii SspB Protein
Article Snippet: Paragraph title: RT-PCR. ... Annealing of the primer and RNA was carried out at 72°C for 2 min and then at 48°C for 1 h. The resulting cDNA was amplified, with each 100 μl of PCR mixture containing 1× PCR buffer, 3 μl of cDNA, 1.5 mM MgCl2 , 10 mM deoxynucleoside triphosphate, 100 ng of each primer, and 2.5 U of Herculase DNA polymerase (Stratagene).

Generated:

Article Title: Chlamydia trachomatis and Chlamydia muridarum spectinomycin resistant vectors and a transcriptional fluorescent reporter to monitor conversion from replicative to infectious bacteria
Article Snippet: PCR was performed using Herculase DNA polymerase (Stratagene). .. Plasmid maps were generated using Serial Cloner ( http://serialbasics.free.fr/Serial_Cloner.html ).

Sequencing:

Article Title: STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites
Article Snippet: PCR was performed using Herculase DNA polymerase (Stratagene). .. PCR primers were obtained from Integrated DNA Technologies and their sequence is listed in . mCh-STIM1 (kind gift from R. Lewis, Stanford, CA) and all truncated and internal deletion were cloned into the EcoR I and Xho I restriction sites of pCDNA3.1+.

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene. .. All wild-type and mutant Utp25 sequences were confirmed by sequence analysis.

Affinity Purification:

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol. .. PCR products were affinity purified with the DNA Clean & Concentrator kit (Zymo Research).

Mutagenesis:

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: Paragraph title: M13KO7 Helper Phage Genome Mutagenesis ... A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol.

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: Paragraph title: Cloning and mutagenesis ... UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene.

Isolation:

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol. .. Purified PCR product was subjected to NheI endonuclease digestion (at the restriction site underlined above), purified by agarose gel electrophoresis, and isolated with a Zymoclean Gel DNA Recovery kit (Zymo Research).

Labeling:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: .. The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, NJ).

Article Title: Spo0A-Dependent Activation of an Extended -10 Region Promoter in Bacillus subtilis
Article Snippet: .. The purified labeled probe was used in a PCR containing 36 pmol of unlabeled SKF4Rev primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, N.J.).

Purification:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: .. The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, NJ).

Article Title: Spo0A-Dependent Activation of an Extended -10 Region Promoter in Bacillus subtilis
Article Snippet: .. The purified labeled probe was used in a PCR containing 36 pmol of unlabeled SKF4Rev primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, N.J.).

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol. .. Purified PCR product was subjected to NheI endonuclease digestion (at the restriction site underlined above), purified by agarose gel electrophoresis, and isolated with a Zymoclean Gel DNA Recovery kit (Zymo Research).

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). .. The PCR product was agarose gel purified using a Qiaquick gel extraction kit, and a series of ligation reactions using increasing amounts of the eluted PCR product were set up overnight at 14 °C.

Polymerase Chain Reaction:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: .. The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, NJ).

Article Title: Chlamydia trachomatis and Chlamydia muridarum spectinomycin resistant vectors and a transcriptional fluorescent reporter to monitor conversion from replicative to infectious bacteria
Article Snippet: .. PCR was performed using Herculase DNA polymerase (Stratagene). .. PCR primers were obtained from Integrated DNA Technologies and are listed in .

Article Title: Spo0A-Dependent Activation of an Extended -10 Region Promoter in Bacillus subtilis
Article Snippet: .. The purified labeled probe was used in a PCR containing 36 pmol of unlabeled SKF4Rev primer and Herculase DNA polymerase (Stratagene). .. The PCR product was purified by elution from a G-50 MicroSpin column (Amersham Pharmacia Biotech, Piscataway, N.J.).

Article Title: H2A.B facilitates transcription elongation at methylated CpG loci
Article Snippet: .. For region C of Kcnq1 , Herculase DNA polymerase (Stratagene) was used for the PCR, with 1× Herculase buffer, 0.3 μM of each primer, 0.8 mM deoxynucleoside triphosphates, 4% dimethyl sulfoxide, and 0.1 μCi of [32 P]dCTP. .. The PCR products were sequenced, and single-nucleotide polymorphism was used to determine the allele-specific deposition of H2A.B.

Article Title: Identification of a Porphyromonas gingivalis Receptor for the Streptococcus gordonii SspB Protein
Article Snippet: .. Annealing of the primer and RNA was carried out at 72°C for 2 min and then at 48°C for 1 h. The resulting cDNA was amplified, with each 100 μl of PCR mixture containing 1× PCR buffer, 3 μl of cDNA, 1.5 mM MgCl2 , 10 mM deoxynucleoside triphosphate, 100 ng of each primer, and 2.5 U of Herculase DNA polymerase (Stratagene). .. To identify molecules of P. gingivalis that interact with SspB protein, surface proteins were first labeled with biotin and extracted with EDTA and mild sonication.

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: .. A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol. .. PCR products were affinity purified with the DNA Clean & Concentrator kit (Zymo Research).

Article Title: A Co-infection Model System and the Use of Chimeric Proteins to Study Chlamydia Inclusion Proteins Interaction
Article Snippet: .. PCR was performed using Herculase DNA polymerase (Stratagene). .. PCR primers were obtained from Integrated DNA Technologies.

Article Title: STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites
Article Snippet: .. PCR was performed using Herculase DNA polymerase (Stratagene). .. PCR primers were obtained from Integrated DNA Technologies and their sequence is listed in . mCh-STIM1 (kind gift from R. Lewis, Stanford, CA) and all truncated and internal deletion were cloned into the EcoR I and Xho I restriction sites of pCDNA3.1+.

Article Title: hERG1 channels are overexpressed in glioblastoma multiforme and modulate VEGF secretion in glioblastoma cell lines
Article Snippet: .. Total cDNA was used as a template for PCR amplification with Herculase DNApol (Stratagene, Cedar Creek, TX) and each of the primer pairs reported below. .. As a quality control on cDNA, PCR amplification of the messenger relative to the housekeeping gene gapdh was performed using the following primer pair: Fw: 5′-AACAGCCTCAAGATCATCAGCAA-3′ Rev: 5′-CAGTCTGGGTGGCAGTGAT-3′ (NG 003027, nucleotides 457–564) Samples positive to gapdh amplification were further analysed using the following primers: hERG1 fw: 5′-TCCAGCGGCTGTACTCGGGC-3′ hERG1 rev: 5′-TGGACCAGAAGTGGTCGGAGAACTC-3′ (U04270, nucleotides 2171–2746). helk2 fw: 5′-CTGCCCTGCGGGGCACTGTCTG-3′ helk2 rev: 5′-AGATCTGGGGGCACATTCCATG-3′ (NM012284 nucleotides 1802–2516).

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: .. The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). ..

Article Title: The Transcriptional Status but Not the Imprinting Control Region Determines Allele-Specific Histone Modifications at the Imprinted H19 Locus ▿
Article Snippet: .. For KvDMR1, Herculase DNA polymerase (Stratagene) was used for the PCR, with 1× Herculase buffer, 0.3 μM of each primer, 0.8 mM deoxynucleoside triphosphates, 4% dimethyl sulfoxide, and 0.1 μCi of [32 P]dCTP. ..

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: .. UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene. .. PCR products were cloned into pCR4-TOPO (Invitrogen) and subsequently subcloned into the NcoI and XhoI sites of p415-GPD (a low-copy yeast expression vector with a constitutive GPD promoter; AmpR , LEU2 marker, and CEN/ARS origin of replication) ( ).

Chromatin Immunoprecipitation:

Article Title: H2A.B facilitates transcription elongation at methylated CpG loci
Article Snippet: Genomic DNA associated with H2A.B was collected by ChIP and amplified by PCR. .. For region C of Kcnq1 , Herculase DNA polymerase (Stratagene) was used for the PCR, with 1× Herculase buffer, 0.3 μM of each primer, 0.8 mM deoxynucleoside triphosphates, 4% dimethyl sulfoxide, and 0.1 μCi of [32 P]dCTP.

Article Title: The Transcriptional Status but Not the Imprinting Control Region Determines Allele-Specific Histone Modifications at the Imprinted H19 Locus ▿
Article Snippet: Paragraph title: Allele-specific ChIP PCRs. ... For KvDMR1, Herculase DNA polymerase (Stratagene) was used for the PCR, with 1× Herculase buffer, 0.3 μM of each primer, 0.8 mM deoxynucleoside triphosphates, 4% dimethyl sulfoxide, and 0.1 μCi of [32 P]dCTP.

Plasmid Preparation:

Article Title: Chlamydia trachomatis and Chlamydia muridarum spectinomycin resistant vectors and a transcriptional fluorescent reporter to monitor conversion from replicative to infectious bacteria
Article Snippet: Paragraph title: Plasmid construction ... PCR was performed using Herculase DNA polymerase (Stratagene).

Article Title: A Co-infection Model System and the Use of Chimeric Proteins to Study Chlamydia Inclusion Proteins Interaction
Article Snippet: Paragraph title: Plasmid construction ... PCR was performed using Herculase DNA polymerase (Stratagene).

Article Title: STIM1 Is a Novel Component of ER-Chlamydia trachomatis Inclusion Membrane Contact Sites
Article Snippet: Paragraph title: Plasmid construction ... PCR was performed using Herculase DNA polymerase (Stratagene).

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: .. The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). ..

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene. .. PCR products were cloned into pCR4-TOPO (Invitrogen) and subsequently subcloned into the NcoI and XhoI sites of p415-GPD (a low-copy yeast expression vector with a constitutive GPD promoter; AmpR , LEU2 marker, and CEN/ARS origin of replication) ( ).

Agarose Gel Electrophoresis:

Article Title: Chemical and Genetic Wrappers for Improved Phage and RNA Display
Article Snippet: A PCR (50 µL reaction) with Herculase™ DNA polymerase (Stratagene) using single-stranded M13KO7 DNA as template and the KO7-P8-AKAS-fwd/rev primer pair was prepared and thermocycled according to the manufacturer’s protocol. .. Purified PCR product was subjected to NheI endonuclease digestion (at the restriction site underlined above), purified by agarose gel electrophoresis, and isolated with a Zymoclean Gel DNA Recovery kit (Zymo Research).

Article Title: hERG1 channels are overexpressed in glioblastoma multiforme and modulate VEGF secretion in glioblastoma cell lines
Article Snippet: Total RNA was quantified by spectrophotometric analysis and checked for purity and quality by running a small aliquot on a 1% agarose gel. .. Total cDNA was used as a template for PCR amplification with Herculase DNApol (Stratagene, Cedar Creek, TX) and each of the primer pairs reported below.

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). .. The PCR product was agarose gel purified using a Qiaquick gel extraction kit, and a series of ligation reactions using increasing amounts of the eluted PCR product were set up overnight at 14 °C.

In Vitro:

Article Title: Promoter Activation by Repositioning of RNA Polymerase
Article Snippet: Paragraph title: Protein preparation and in vitro footprinting assays. ... The purified labeled probe was used in a PCR mixture containing 36 pmol of unlabeled primer and Herculase DNA polymerase (Stratagene).

Marker:

Article Title: The DEAD-box RNA helicase-like Utp25 is an SSU processome component
Article Snippet: UTP25 was PCR-amplified from yeast total DNA (YPH499) using Herculase DNA polymerase (Stratagene) and oligonucleotide primers that incorporate NcoI and XhoI restriction sites into the 5′- and 3′-ends of the gene. .. PCR products were cloned into pCR4-TOPO (Invitrogen) and subsequently subcloned into the NcoI and XhoI sites of p415-GPD (a low-copy yeast expression vector with a constitutive GPD promoter; AmpR , LEU2 marker, and CEN/ARS origin of replication) ( ).

Gel Extraction:

Article Title: Kinetic modification of the ?1I subunit-mediated T-type Ca2+ channel by a human neuronal Ca2+ channel ? subunit
Article Snippet: The PCR components were 50 ng plasmid DNA, 100 ng each primer, 0.2 m m dNTPs, 1 × Herculase reaction buffer and 2.5 U Herculase DNA polymerase (Stratagene). .. The PCR product was agarose gel purified using a Qiaquick gel extraction kit, and a series of ligation reactions using increasing amounts of the eluted PCR product were set up overnight at 14 °C.

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  • 87
    Stratagene herculase ii fusion dna polymerase pcr system
    Analysis of replication timing changes after the insertion of cis -regulatory elements. (A) Diagram showing the modified allele after insertion of a transgene by homologous recombination and below the wild type allele. Primer pairs used for replication timing analyses are shown and can detect replication timing of the modified allele (With), the wild type allele (Without), or both alleles (Both). (B) After insertion by homologous recombination, selected clones are pulse labeled with BrdU and cell sorted into four fractions from early to late S-phase. BrdU labeled nascent strand <t>DNA</t> is immunoprecipitated and quantified by real-time <t>PCR.</t> The nascent strands produced at the site of integration on the allele containing the transgene (With), the wild type allele (Without), or both alleles (Both) were quantified. Analyses of both alleles should display an average picture of the profiles obtained on each allele, thus providing a validation of the PCR quantification. For each clone, we calculate differences in the timing of replication between the unmodified (Without) and targeted (With) alleles, where ΔL and ΔE describe the differences in late and early S-phase, respectively.
    Herculase Ii Fusion Dna Polymerase Pcr System, supplied by Stratagene, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/herculase ii fusion dna polymerase pcr system/product/Stratagene
    Average 87 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    herculase ii fusion dna polymerase pcr system - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    93
    Stratagene herculase ii dna polymerase
    Translation of a <t>DNA</t> template into PNA and displacement of the resulting PNA strand Following PNA polymerization (lane 5) under conditions described in Figure 2 , the PNA strand is displaced by primer extension of the 3′ hairpin using <t>Herculase</t> DNA polymerase II and dNTPs (lane 6). DNA polymerization to the end of the template strand (through the 5′ stem-loop) generates an 84-base pair DNA duplex that can form as a mutually exclusive alternative to the 40-base pair PNA-DNA heteroduplex and 24-base pair DNA-DNA stem. Successful strand displacement creates a double-stranded Bcc I restriction endonuclease cleavage site. The Bcc I-mediated cleavage of the primer extension product (lanes 4 and 8) is therefore consistent with displacement of the PNA strand. The red dot represents a Cy3 backbone spacer that blocks subsequentual DNA polymerization. The green dot represents a fluorescein group. The denaturing PAGE gel shown is visualized by imaging fluorescein fluorescence. Reactions shown in lanes labeled “translation–” lacked NaCNBH 3 ; those in lanes labeled “displacement–” lacked DNA polymerase; those in lanes labeled “restriction digestion–” lacked Bcc I.
    Herculase Ii Dna Polymerase, supplied by Stratagene, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/herculase ii dna polymerase/product/Stratagene
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    herculase ii dna polymerase - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    94
    Stratagene long range high fidelity amplification dna polymerase
    Schematic organization of JcDV-based plasmids used to generate linear sequences for transfection experiments. pBR322 backbone is figured as dotted gray line. JcDV structural proteins ( VP ) coding sequences are represented by a solid black line; non-structural proteins ( NS ) genes by a solid gray line. Both share a polyadenylation signal shown as an open ellipse. Open boxes figure the <t>p9</t> and p93 ITRs, hatched boxes underline the location of p9 and p93 promoters, respectively. GFP coding sequence is figured with a gray box and its 3′ SV40-derived polyadenylation signal is shown as a hatched line. Arrows numbered according to Table 1 figure the primers used for PCR-based experiments. Primers used for walk-PCR are represented above; primers 7–13 represented only relatively to p9 can also hybridize to p93 <t>DNA</t> sequences when present. Gray-filled arrowheads figure restriction sites used to generate linear molecules from the plasmid constructs. Open arrowheads figure restriction sites used for walk-PCR experiments. Subscript numbers indicate iterated restriction sites. By convention, nucleotide numbers of each linear molecule are accorded to the 5′ C generated after Cla I restriction (AT/CGAT). Restriction enzymes are: A: Afl II, C: Cla I, D: Dra I, H: Hpa I, M: Msp A1L, S: Ssp I, P: Psh AI, Pv: Pvu II, X: Xcm I, respectively. Linear molecules were obtained after restriction of three different JcDV-based vectors. Their length is indicated under the name of each plasmid: (A) pFull encompassing a full-length sequence of JcDV DNA and the GFP marker gene, cloned into pBR322 plasmid. This schematic representation displays all the symbols described above; some of them only are reported in B and C. (B) pVP in comparison to pFull, a frameshift deletion affects the NS region (C) pNS in comparison to pFull, lacks VP genes. The expression of GFP is directly under the control of the p9 promoter. Primers giving rise to specific products after PCR are shown (See Table 1 ).
    Long Range High Fidelity Amplification Dna Polymerase, supplied by Stratagene, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/long range high fidelity amplification dna polymerase/product/Stratagene
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    long range high fidelity amplification dna polymerase - by Bioz Stars, 2020-04
    94/100 stars
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    Analysis of replication timing changes after the insertion of cis -regulatory elements. (A) Diagram showing the modified allele after insertion of a transgene by homologous recombination and below the wild type allele. Primer pairs used for replication timing analyses are shown and can detect replication timing of the modified allele (With), the wild type allele (Without), or both alleles (Both). (B) After insertion by homologous recombination, selected clones are pulse labeled with BrdU and cell sorted into four fractions from early to late S-phase. BrdU labeled nascent strand DNA is immunoprecipitated and quantified by real-time PCR. The nascent strands produced at the site of integration on the allele containing the transgene (With), the wild type allele (Without), or both alleles (Both) were quantified. Analyses of both alleles should display an average picture of the profiles obtained on each allele, thus providing a validation of the PCR quantification. For each clone, we calculate differences in the timing of replication between the unmodified (Without) and targeted (With) alleles, where ΔL and ΔE describe the differences in late and early S-phase, respectively.

    Journal: PLoS Biology

    Article Title: USF Binding Sequences from the HS4 Insulator Element Impose Early Replication Timing on a Vertebrate Replicator

    doi: 10.1371/journal.pbio.1001277

    Figure Lengend Snippet: Analysis of replication timing changes after the insertion of cis -regulatory elements. (A) Diagram showing the modified allele after insertion of a transgene by homologous recombination and below the wild type allele. Primer pairs used for replication timing analyses are shown and can detect replication timing of the modified allele (With), the wild type allele (Without), or both alleles (Both). (B) After insertion by homologous recombination, selected clones are pulse labeled with BrdU and cell sorted into four fractions from early to late S-phase. BrdU labeled nascent strand DNA is immunoprecipitated and quantified by real-time PCR. The nascent strands produced at the site of integration on the allele containing the transgene (With), the wild type allele (Without), or both alleles (Both) were quantified. Analyses of both alleles should display an average picture of the profiles obtained on each allele, thus providing a validation of the PCR quantification. For each clone, we calculate differences in the timing of replication between the unmodified (Without) and targeted (With) alleles, where ΔL and ΔE describe the differences in late and early S-phase, respectively.

    Article Snippet: We used Herculase II Fusion DNA Polymerase PCR system (Stratagene) for the amplification with the following conditions: the initial denaturation at 95°C for 2 min, and 35 cycles of 95°C for 30 s, 57°C for 30 s, and 72°C for 3 min and a final extension of 72°C for 5 min. At first, the PCR product was cloned with Gateway BP Clonase system (Invitrogen) to generate the entry vector.

    Techniques: Modification, Homologous Recombination, Clone Assay, Labeling, Immunoprecipitation, Real-time Polymerase Chain Reaction, Produced, Polymerase Chain Reaction

    Translation of a DNA template into PNA and displacement of the resulting PNA strand Following PNA polymerization (lane 5) under conditions described in Figure 2 , the PNA strand is displaced by primer extension of the 3′ hairpin using Herculase DNA polymerase II and dNTPs (lane 6). DNA polymerization to the end of the template strand (through the 5′ stem-loop) generates an 84-base pair DNA duplex that can form as a mutually exclusive alternative to the 40-base pair PNA-DNA heteroduplex and 24-base pair DNA-DNA stem. Successful strand displacement creates a double-stranded Bcc I restriction endonuclease cleavage site. The Bcc I-mediated cleavage of the primer extension product (lanes 4 and 8) is therefore consistent with displacement of the PNA strand. The red dot represents a Cy3 backbone spacer that blocks subsequentual DNA polymerization. The green dot represents a fluorescein group. The denaturing PAGE gel shown is visualized by imaging fluorescein fluorescence. Reactions shown in lanes labeled “translation–” lacked NaCNBH 3 ; those in lanes labeled “displacement–” lacked DNA polymerase; those in lanes labeled “restriction digestion–” lacked Bcc I.

    Journal: Nature chemical biology

    Article Title: An In Vitro Translation, Selection, and Amplification System for Peptide Nucleic Acids

    doi: 10.1038/nchembio.280

    Figure Lengend Snippet: Translation of a DNA template into PNA and displacement of the resulting PNA strand Following PNA polymerization (lane 5) under conditions described in Figure 2 , the PNA strand is displaced by primer extension of the 3′ hairpin using Herculase DNA polymerase II and dNTPs (lane 6). DNA polymerization to the end of the template strand (through the 5′ stem-loop) generates an 84-base pair DNA duplex that can form as a mutually exclusive alternative to the 40-base pair PNA-DNA heteroduplex and 24-base pair DNA-DNA stem. Successful strand displacement creates a double-stranded Bcc I restriction endonuclease cleavage site. The Bcc I-mediated cleavage of the primer extension product (lanes 4 and 8) is therefore consistent with displacement of the PNA strand. The red dot represents a Cy3 backbone spacer that blocks subsequentual DNA polymerization. The green dot represents a fluorescein group. The denaturing PAGE gel shown is visualized by imaging fluorescein fluorescence. Reactions shown in lanes labeled “translation–” lacked NaCNBH 3 ; those in lanes labeled “displacement–” lacked DNA polymerase; those in lanes labeled “restriction digestion–” lacked Bcc I.

    Article Snippet: Herculase II DNA polymerase (Stratagene), a fusion protein of Pfu Ultra and a DNA-binding domain that is designed to facilitate DNA polymerization on GC-rich templates, was able to displace the PNA strand efficiently ( ).

    Techniques: Polyacrylamide Gel Electrophoresis, Imaging, Fluorescence, Labeling

    Schematic organization of JcDV-based plasmids used to generate linear sequences for transfection experiments. pBR322 backbone is figured as dotted gray line. JcDV structural proteins ( VP ) coding sequences are represented by a solid black line; non-structural proteins ( NS ) genes by a solid gray line. Both share a polyadenylation signal shown as an open ellipse. Open boxes figure the p9 and p93 ITRs, hatched boxes underline the location of p9 and p93 promoters, respectively. GFP coding sequence is figured with a gray box and its 3′ SV40-derived polyadenylation signal is shown as a hatched line. Arrows numbered according to Table 1 figure the primers used for PCR-based experiments. Primers used for walk-PCR are represented above; primers 7–13 represented only relatively to p9 can also hybridize to p93 DNA sequences when present. Gray-filled arrowheads figure restriction sites used to generate linear molecules from the plasmid constructs. Open arrowheads figure restriction sites used for walk-PCR experiments. Subscript numbers indicate iterated restriction sites. By convention, nucleotide numbers of each linear molecule are accorded to the 5′ C generated after Cla I restriction (AT/CGAT). Restriction enzymes are: A: Afl II, C: Cla I, D: Dra I, H: Hpa I, M: Msp A1L, S: Ssp I, P: Psh AI, Pv: Pvu II, X: Xcm I, respectively. Linear molecules were obtained after restriction of three different JcDV-based vectors. Their length is indicated under the name of each plasmid: (A) pFull encompassing a full-length sequence of JcDV DNA and the GFP marker gene, cloned into pBR322 plasmid. This schematic representation displays all the symbols described above; some of them only are reported in B and C. (B) pVP in comparison to pFull, a frameshift deletion affects the NS region (C) pNS in comparison to pFull, lacks VP genes. The expression of GFP is directly under the control of the p9 promoter. Primers giving rise to specific products after PCR are shown (See Table 1 ).

    Journal: PeerJ

    Article Title: Linear Lepidopteran ambidensovirus 1 sequences drive random integration of a reporter gene in transfected Spodoptera frugiperda cells

    doi: 10.7717/peerj.4860

    Figure Lengend Snippet: Schematic organization of JcDV-based plasmids used to generate linear sequences for transfection experiments. pBR322 backbone is figured as dotted gray line. JcDV structural proteins ( VP ) coding sequences are represented by a solid black line; non-structural proteins ( NS ) genes by a solid gray line. Both share a polyadenylation signal shown as an open ellipse. Open boxes figure the p9 and p93 ITRs, hatched boxes underline the location of p9 and p93 promoters, respectively. GFP coding sequence is figured with a gray box and its 3′ SV40-derived polyadenylation signal is shown as a hatched line. Arrows numbered according to Table 1 figure the primers used for PCR-based experiments. Primers used for walk-PCR are represented above; primers 7–13 represented only relatively to p9 can also hybridize to p93 DNA sequences when present. Gray-filled arrowheads figure restriction sites used to generate linear molecules from the plasmid constructs. Open arrowheads figure restriction sites used for walk-PCR experiments. Subscript numbers indicate iterated restriction sites. By convention, nucleotide numbers of each linear molecule are accorded to the 5′ C generated after Cla I restriction (AT/CGAT). Restriction enzymes are: A: Afl II, C: Cla I, D: Dra I, H: Hpa I, M: Msp A1L, S: Ssp I, P: Psh AI, Pv: Pvu II, X: Xcm I, respectively. Linear molecules were obtained after restriction of three different JcDV-based vectors. Their length is indicated under the name of each plasmid: (A) pFull encompassing a full-length sequence of JcDV DNA and the GFP marker gene, cloned into pBR322 plasmid. This schematic representation displays all the symbols described above; some of them only are reported in B and C. (B) pVP in comparison to pFull, a frameshift deletion affects the NS region (C) pNS in comparison to pFull, lacks VP genes. The expression of GFP is directly under the control of the p9 promoter. Primers giving rise to specific products after PCR are shown (See Table 1 ).

    Article Snippet: The long p9 ITR-NS region was assessed with primers located in the p9 ITR (1′) and the NS-3 (4), using a long range/high fidelity amplification DNA polymerase (Herculase; Stratagene Agilent Technologies, Les Ulis, France).

    Techniques: Transfection, Sequencing, Derivative Assay, Polymerase Chain Reaction, Plasmid Preparation, Construct, Generated, Marker, Clone Assay, Expressing

    Schematic representation of viral and genomic DNA sequences identified in our experiments. Raw sequences obtained after numerous amplifications of viral-genomic junctions ( Supplemental Information ) where aligned against (i) the sequence of the transfected linear plasmid using the discontiguous megablast conditions, (ii) the nr/nt nucleotide collection specified to Spodoptera frugiperda (taxid 7108), using the blastn conditions. Only the cleaned sequences are represented and motifs identify contiguous sequences from the same amplified sequence, named on the right. (A) BLAST-based alignment against lpFull of raw sequences obtained after amplifications of viral junctions from lpFull-transfected cells. The open diamond represents the location of ∼20 nts from the Bac 68E14 from S. frugiperda [id:681381.1] as a significative example of the rearrangement. (B) BLAST-based alignment against lpVP of raw sequences obtained after amplifications of viral junctions from lpVP-transfected cells. The triangle represents 70 nts of BAC 75E05 from S. frugiperda [id: 681368.1] flanking a 40 nts fragment of the ITR (2–40) and the ellipse represents 70 nts of the same BAC intercalated between a JcDV VP fragment (3,829–3,531) and the NS -ITR boundary (771–551). (C) BLAST alignment against lpNS of raw sequences obtained after amplifications of viral junctions from lpNS-transfected cells.

    Journal: PeerJ

    Article Title: Linear Lepidopteran ambidensovirus 1 sequences drive random integration of a reporter gene in transfected Spodoptera frugiperda cells

    doi: 10.7717/peerj.4860

    Figure Lengend Snippet: Schematic representation of viral and genomic DNA sequences identified in our experiments. Raw sequences obtained after numerous amplifications of viral-genomic junctions ( Supplemental Information ) where aligned against (i) the sequence of the transfected linear plasmid using the discontiguous megablast conditions, (ii) the nr/nt nucleotide collection specified to Spodoptera frugiperda (taxid 7108), using the blastn conditions. Only the cleaned sequences are represented and motifs identify contiguous sequences from the same amplified sequence, named on the right. (A) BLAST-based alignment against lpFull of raw sequences obtained after amplifications of viral junctions from lpFull-transfected cells. The open diamond represents the location of ∼20 nts from the Bac 68E14 from S. frugiperda [id:681381.1] as a significative example of the rearrangement. (B) BLAST-based alignment against lpVP of raw sequences obtained after amplifications of viral junctions from lpVP-transfected cells. The triangle represents 70 nts of BAC 75E05 from S. frugiperda [id: 681368.1] flanking a 40 nts fragment of the ITR (2–40) and the ellipse represents 70 nts of the same BAC intercalated between a JcDV VP fragment (3,829–3,531) and the NS -ITR boundary (771–551). (C) BLAST alignment against lpNS of raw sequences obtained after amplifications of viral junctions from lpNS-transfected cells.

    Article Snippet: The long p9 ITR-NS region was assessed with primers located in the p9 ITR (1′) and the NS-3 (4), using a long range/high fidelity amplification DNA polymerase (Herculase; Stratagene Agilent Technologies, Les Ulis, France).

    Techniques: Sequencing, Transfection, Plasmid Preparation, Amplification, BAC Assay

    NS genes sequences are highly rearranged in stable fluorescent cell clones. (A) PCR amplification of the VP -GFP region with primers one and two (see Table 1 ; Fig. 1 ) from lpFull and lpVP clones. (B) Amplification of the large p9 ITR- NS region using primers pair 1′, 4. (C) Amplification of the NS -GFP region using primers pair 3, 4. (D) Amplification of ORF2, ORF3 and ORF4 RNA transcripts by RT-PCR using primers in the untranslated region (UTR) upstream the NS-3 ATG codon and the NS-1 region (see Table 1 ). Molecular weight marker (Mk) band sizes are shown on each gel. pFull was used as positive control. Sf9 DNA was used as a negative control.

    Journal: PeerJ

    Article Title: Linear Lepidopteran ambidensovirus 1 sequences drive random integration of a reporter gene in transfected Spodoptera frugiperda cells

    doi: 10.7717/peerj.4860

    Figure Lengend Snippet: NS genes sequences are highly rearranged in stable fluorescent cell clones. (A) PCR amplification of the VP -GFP region with primers one and two (see Table 1 ; Fig. 1 ) from lpFull and lpVP clones. (B) Amplification of the large p9 ITR- NS region using primers pair 1′, 4. (C) Amplification of the NS -GFP region using primers pair 3, 4. (D) Amplification of ORF2, ORF3 and ORF4 RNA transcripts by RT-PCR using primers in the untranslated region (UTR) upstream the NS-3 ATG codon and the NS-1 region (see Table 1 ). Molecular weight marker (Mk) band sizes are shown on each gel. pFull was used as positive control. Sf9 DNA was used as a negative control.

    Article Snippet: The long p9 ITR-NS region was assessed with primers located in the p9 ITR (1′) and the NS-3 (4), using a long range/high fidelity amplification DNA polymerase (Herculase; Stratagene Agilent Technologies, Les Ulis, France).

    Techniques: Polymerase Chain Reaction, Amplification, Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Marker, Positive Control, Negative Control