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Herculase dna polymerase

Manufactured by Agilent Technologies
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Herculase DNA polymerase is a high-fidelity DNA polymerase used for PCR amplification of DNA. It has a 3' to 5' exonuclease proofreading activity, which results in a low error rate during DNA synthesis.

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Lab products found in correlation

Taq dna polymerase, by Takara Bio (1 mentions)

Spelling variants of this product

Herculase II Fusion DNA Polymerase (191 citations) Herculase II DNA polymerase (17 citations) Herculase polymerase (7 citations) Herculase II Fusion DNA Polymerase Kit (6 citations) Herculase II Phusion DNA polymerase (4 citations) High-fidelity Herculase II DNA polymerases (3 citations)

3 protocols using herculase dna polymerase

1

Sperm mtDNA Amplification Protocol

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The desired long segment of targeted sequences of mtDNA was amplified from 100–150 ng of sperm mtDNA in a 50 µL reaction mixture, containing 200 mM of each dNTP (Merck Biosciences, Bangalore), 1 mM of each light and heavy strand primers (IDT Technologies, USA), 1.5 U of herculase DNA polymerase, 50 mM KCL, 2 mM Mgcl2,25 mM Tris[hydromethyl] methyl-3-aminopropanesulphonic acid (Agilent Technologies, USA), 1 mM – mercaptoethanol, and 10 mM Tris-HCL (pH 8.5). PCR was carried out for 25 cycles in two steps in first 12 cycles using the denaturation at 94°C for 1 min, annealing at 55–58°C for 1 min, and primer extension at 72°C for 6 min and remaining 13 cycles denaturation and annealing condition were same but primer extension at 68°C for increasing 10 s to each reactions.[14 (link)] The long-range PCR products were then separated on a 1% ethidium bromide containing agarose gel for 1.5 h and observed on gel documentation system (Uvi-Teck, UK).
Ambulkar P.S., Chuadhari A.R, & Pal A.K. (2016). Association of large scale 4977-bp “common” deletions in sperm mitochondrial DNA with asthenozoospermia and oligoasthenoteratozoospermia. Journal of Human Reproductive Sciences, 9(1), 35-40.
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2

Molecular Characterization of CDAs

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Standard molecular techniques [74 ] were used to prepare the complementation constructs with fluorescently tagged CDAs. A set of transformation vectors based on pUCAP was generated as described in Samalova et al. [75 (link)]. The vectors contain polyadenylation signal pATrpC and either bialophos or hygromycin resistance marker that was cloned into re-created SalI sites using primer pairs 1/2 or 3/4 respectively (see S2 Table and S12 Fig). For PCR amplification of CBP1 and CDA2, primer pairs 27/28 and 29/30 were used, respectively. Genomic DNA from the WT strain Guy11 was used as a template, and amplified using Herculase DNA polymerase (Agilent). This resulted in amplification of the coding sequence of the genes (without stop codons), together with 2 kb of native promoter sequence for CBP1 and 1.3 kb for CDA2. The PCR products were cloned into the AscI sites of the vector described above (S12 Fig), creating C-terminal mCherry fusions.
Geoghegan I.A, & Gurr S.J. (2016). Chitosan Mediates Germling Adhesion in Magnaporthe oryzae and Is Required for Surface Sensing and Germling Morphogenesis. PLoS Pathogens, 12(6), e1005703.
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3

Amplification and Sequencing of Ig Introns

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Intron sequences were amplified using Herculase DNA polymerase (Agilent) and nested primers for V genes and intronic enhancers (Table SI). PCR products were dA-tailed using Taq DNA polymerase (Takara) and cloned into StrataClone pSC-Amp/Kan vector (Agilent). During amplification, the smaller J-intron (JH3, JH4, Jκ4, and Jκ5) sequences were preferentially amplified. They were then extracted from the gel and cloned for sequencing. DNA from the larger PCR products (JH1, JH2, Jκ1, Jκ2) were obtained from total amplified DNA that was cloned and expressed in E. coli. Colonies were screened using 32P-end-labeled oligonucleotides specific for the intron sequences, which bound downstream of each J gene (Table SI). Plasmid inserts were sequenced by Sanger sequencing; only clones with unique VDJ and VJ joins were examined. The data was then compared to the C57BL/6 genomic sequence to identify mutations. Mutations were counted at the beginning of the J intron to avoid selection bias in the V(D)J exon. Mutational background from PCR and Sanger sequencing was previously measured as 1.8 x 10−4 mutations/bp (20 (link)). Sequences are available on request.
Heltzel J.H., Maul R.W., Yang W, & Gearhart P.J. (2022). Promoter Proximity Defines Mutation Window for VH and VK Genes Rearranged to Different J Genes. Journal of immunology (Baltimore, Md. : 1950), 208(9), 2220-2226.
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