Herculase 2 fusion dna polymerase kit

Manufactured by Agilent Technologies

Sourced in United States

The Herculase II Fusion DNA Polymerase Kit is a laboratory reagent designed for high-fidelity DNA amplification. It is a DNA polymerase enzyme that can be used for a variety of molecular biology applications requiring accurate DNA replication.

Automatically generated - may contain errors

Lab products found in correlation

6 protocols using herculase 2 fusion dna polymerase kit

Nested PCR Amplification of Gene Sequence

Check if the same lab product or an alternative is used in the 5 most similar protocols

For amplification a nested PCR approach was chosen. The Herculase II Fusion DNA Polymerase Kit (Agilent Technologies) was used in a 25 μL reaction volume. 24 μL of Master Mix were mixed with 1 μL of extracted patient gDNA or purified first PCR. Each PCR was carried out in triplicates to normalize for PCR amplification bias. First PCR included sense primer F-6553 5′- ATGGGATCAAAGCCTAAAGCCATGTG-3′ and R-7801 5′- AGTGCTTCCTGCTGCTCCCAAGAACCCAAG-3′. Following cycling conditions were used: initial 3 min at 95°C, 30 cycles of denaturation for 15 s at 95°C, annealing for 20 s at 60°C and extension for 45 s at 72°C. Final extension was done for 3 min at 72°C.
The second PCR included sense primer F-6848 (5′- AGGCCTGTCCAAAGGTATCCTTTGA-3′; second PCR) and antisense primer R- 7371 (5′- TTACAGTAGAAAAATTCCCCTCCACAATTAAA-3′; second PCR). Cycling conditions for the second PCR were the same as for the first PCR except an annealing temperature of 56°C for 20 s.

Otte F., Zhang Y., Spagnuolo J., Thielen A., Däumer M., Wiethe C., Stoeckle M., Kusejko K., Klein F., Metzner K.J, & Klimkait T. (2023). Revealing viral and cellular dynamics of HIV-1 at the single-cell level during early treatment periods. Cell Reports Methods, 3(6), 100485.

+ Open protocol

+ Expand

Targeted Sequencing Library Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following materials were used in this study: QIAGEN QlAampDNA Mini Kit (Qiagen, Valencia, Germany); QIAGEN QlAampBlood Mini Kit (Qiagen, Valencia, Germany); Ion Xpress Plus Fragment Library Kit (Life Technologies, New York, USA); Agencourt AMPure XP Kit (Beckman, USA); H2O (Sigma-Aldrich, USA); Thermo NanoDrop 1000 (Thermo, MA, USA); SureSelect Target Enrichment System for Sequencing on Ion-Proton (Agilent Technologies, USA) ; SureSelect TE Reagent Kit, PTN (Agilent Technologies, USA); Herculase II Fusion DNA Polymerase kit (Agilent Technologies, USA); Ion OneTouch 2 System (Life Technologies, New York, USA); Qubit 2.0 (Life Technologies, New York, USA); Ion PI Template OT2 200 Kit v2 (Life Technologies, New York, USA); Ion PI Sequencing 200 Kit v2 (Life Technologies, New York, USA); ABI 3730xl Sequencing Machine (Life Technologies, New York, USA); and PCR machine (Life Technologies, New York, USA); QIAxcel Bioanalyzer (Qiagen, Valencia, Germany); ABI Ion-Proton Sequencing Machine (Life Technologies, New York, USA).

Chen Y., Zhang X.C., Yan W.Q., Guo W.B., Xie Z., Lu D.X., Lv Z.Y., Chen Z.H, & Su J. (2020). Establishment and application of a method of next generation sequencing of 285 genes in lung cancer based on Ion-Proton platform. Translational Cancer Research, 9(7), 4239-4249.

+ Open protocol

+ Expand

YN1mAb Hybridoma DNA Extraction and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

The YN1mAb hybridoma genomic DNA was extracted using GeneJet genomic DNA purification kit from Thermo Fisher Scientific (Waltham, MA). PCR analysis was carried out using primers covering the C-terminal end of the YN1mAb. Herculase II Fusion DNA Polymerase Kit from Agilent Technologies (Santa Clara, CA) was used for PCR analysis. The PCR amplicons were run on a 2% (wt/vol) agarose gel and stained with ethidium bromide. Sanger sequencing was carried out for confirmation of the insert.

Khoshnejad M., Brenner J.S., Motley W., Parhiz H., Greineder C.F., Villa C.H., Marcos-Contreras O.A., Tsourkas A, & Muzykantov V.R. (2018). Molecular engineering of antibodies for site-specific covalent conjugation using CRISPR/Cas9. Scientific Reports, 8, 1760.

+ Open protocol

+ Expand

Generation of CVB3 Infectious Clones

Check if the same lab product or an alternative is used in the 5 most similar protocols

To generate a non-reporter wild type infectious clone of CVB3 [strain Woodruff, also known as H3 strain], the total RNA from wild type CVB3 infected cells was extracted. Subsequently, the cDNA was generated using SuperScript^™ IV First-Strand Synthesis System (Invitrogen, Cat#18091050) to be template. Then, we used PCR to clone the full length CVB3 genome using Herculase II Fusion DNA Polymerase kit (Agilent Technologies, Inc., Cat#600675), and cloned it into pCR-BluntII-TOPO vector (Invitrogen, Cat#K280002). This plasmid was named pCRII-T7-CVB3. The constructs were sequenced to verify the original sequence. The 2C mutated CVB3 used in this study was engineered by using overlap PCR to generate a fragment containing the mutation, then inserted into CVB3 infectious cDNA constructs using SpeI and PmlI.
For CVB3 replicon constructs, we modified pCVB3-eGFP infectious clone (Feuer et al., 2002 (link)) by inserting Rluc2A coding sequence using BstBI and NdeI restriction sites. The wild type CVB3 replicon was termed pCVB3-Rluc2A-RepWT. To make a replication-deficient replicon, the codons of the GDD polymerase active site “ggt gac gat” in viral 3D gene were replaced with “acg cgt”, which encodes amino acids TR, and is recognized by restriction enzyme MluI. This plasmid was named pCVB3-RLuc2A-RepMut.

Fan W., Mar K.B., Sari L., Gaszek I.K., Cheng Q., Evers B.M., Shelton J.M., Wight-Carter M., Siegwart D.J., Lin M.M, & Schoggins J.W. (2021). TRIM7 inhibits enterovirus replication and promotes emergence of a viral variant with increased pathogenicity. Cell, 184(13), 3410-3425.e17.

+ Open protocol

+ Expand

Nanopore Sequencing Amplicon Generation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Amplicons for the nanopore sequencing were generated using the Herculase II fusion DNA Polymerase kit (Agilent, Waldbronn, Germany) following the manufacturer’s protocol. The whole genomic region was divided into 10-15 kb fragments, which were amplified in both strains (RS5346 “no ubas” and RSA016 “high ubas”) using specific primers (see Key Resources Table). Primer design was carried out using Geneious software (version 8.0.1). In brief, the conditions for PCR experiment were the following: initial denaturation for 2 minutes at 95 °C, ten cycles of denaturation at 95 °C for 20 seconds, annealing at 60°C for 20 seconds, and extension at 68 °C for 8 min, followed by 20 cycles of denaturation at 95°C for 20 seconds, annealing at 60 °C for 20 seconds, and extension at 68 °C 8 min, plus 20 seconds longer per cycle and a final extension 68°C for 8 minutes.

Falcke J.M., Bose N., Artyukhin A.B., Rödelsperger C., Markov G.V., Yim J.J., Grimm D., Claassen M.H., Panda O., Baccile J.A., Zhang Y.K., Le H.H., Jolic D., Schroeder F.C, & Sommer R.J. (2018). Linking Genomic and Metabolomic Natural Variation Uncovers Nematode Pheromone Biosynthesis. Cell chemical biology, 25(6), 787-796.e12.

+ Open protocol

+ Expand

Genomic DNA Extraction and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissue genomic DNA extraction kit DP304 (TIANGEN), KAPA HyperPlus Kits (Roche), HyperCap Bead Kit (Roche), SureSelect Target Enrichment Kit ILM Indexing Hyb Module Box 2 (Agilent), PlateLoc Thermal Microplate Sealer (Agilent), Herculase II Fusion DNA Polymerase Kit (Agilent), Sequencing and Library Building Platform (IIIumina USA) were used.

Song P., Yang D., Wang H., Cui X., Si X., Zhang X, & Zhang L. (2020). Relationship between the efficacy of immunotherapy and characteristics of specific tumor mutation genes in non‐small cell lung cancer patients. Thoracic Cancer, 11(6), 1647-1654.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!