Primestar kit

Manufactured by Takara Bio

Sourced in Japan, China

The PrimeSTAR Kit is a high-fidelity DNA polymerase system designed for accurate and efficient DNA amplification. It provides reliable performance for a wide range of applications, including gene cloning, site-directed mutagenesis, and long-fragment PCR.

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Lab products found in correlation

Hiscript 2 reverse transcriptase kit, by Vazyme (1 mentions) Rneasy plant mini kit, by Qiagen (1 mentions) Revertra ace kit, by Toyobo (1 mentions) Ddh2o, by Takara Bio (1 mentions) Rnase free h2o, by Takara Bio (1 mentions)

Spelling variants of this product

PrimeSTAR Mutagenesis Basal Kit (188 citations) PrimeSTAR Mutagenesis Kit (20 citations) PrimeSTAR® HS DNA Polymerase kit (13 citations) PrimeSTAR GXL DNA polymerase kit (11 citations) PrimeSTAR Max DNA Polymerase kit (11 citations) PrimeSTAR DNA Polymerase Kit (4 citations) PrimeSTAR max mutagenesis kit (3 citations) PrimeSTAR HS kit (2 citations) PrimeStar® high‐fidelity thermostable DNA polymerase reagent kit (2 citations) PrimeSTAR Max Mutagenesis Basal Kit (2 citations)

5 protocols using primestar kit

Capturing and Characterizing Phosphoglucomutase in Gracilaria lemaneiformis

Cited 1 time

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The whole genome sequences of G. lemaneiformis were captured from our own laboratory database [38 (link)]. For catching the PGM protein sequence, the Hidden Markov Model (HMM) analysis was used for the search. The HMM profiles of the phosphoglucomutase/phosphomannomutase (PF02878, PF02879, PF02880 and PF00408) from the Pfam protein family database Pfam version 35.0 (http://pfam.xfam.org/, 29 June 2022) were used as the query to search the G. lemaneiformis protein database with an e-value ≤ e⁻¹⁰. Blastp was subsequently used to search for missing possible PGM candidates. Pfam and SMART (http://smart.embl-heidelberg.de/, 29 June 2022) were then used to confirm the conserved domain, and the protein sequences lacking the conserved domains were excluded.
The total RNA from G. lemaneiformis was isolated by a RNeasy Plant mini kit (QIAGEN, Germany), according to the manufacturer’s protocol. The cDNA for the full-length sequence cloning was subsequently synthetized by using HiScript II Reverse Transcriptase Kit (Vazyme, Nanjing, China). The opening reading frames (ORFs) of three possible GlPGM were annotated as GlPGM1, GlPGM2 and GlPGM3, and finally amplified by PCR, using Prime Star Kit (Takara, Dalian, China). The gene-specific primers with different restriction endonuclease site are indicated in the additional file Table S1 (Supplementary Materials).

Chen Q., Yu X., Liu S., Luo S., Chen X., Xu N, & Sun X. (2022). Identification, Characteristics and Function of Phosphoglucomutase (PGM) in the Agar Biosynthesis and Carbon Flux in the Agarophyte Gracilariopsis lemaneiformis (Rhodophyta). Marine Drugs, 20(7), 442.

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CRISPR-Cas9 Silencing of Goat miRNA-26 Precursors

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The sgRNAs targeting the genomic sequence of goat pre-miR-26a and pre-miR-26b were designed using the CHOPCHOP website (http://chopchop.cbu.uib.no/, accessed on 9 November 2020). Three sgRNAs with high prediction scores and targeting both ends of the pre-miR-26a and pre-miR-26b sequences were selected, respectively (Figure 1A,B), and synthesized by Sangon Biotech (Shanghai, China) Co., Ltd. Next, the double-stranded oligonucleotides formed by the annealing of sgRNA were inserted into the Bbs I site of plasmid PX459 to construct a vector co-expressing sgRNA and Cas9, named 26a-sg1-PX459, 26a-sg2-PX459, 26a-sg3-PX459, 26b-sg1-PX459, 26b-sg2-PX459, and 26b-sg3-PX459, respectively. U6-26a-sg2-tracRNA, U6-26a-sg3-tracRNA, U6-26b-sg2-tracRNA, and U6-26b-sg3-tracRNA sequences were cloned from 26a-sg2-PX459, 26a-sg3-PX459, 26b-sg2-PX459, and 26b-sg3-PX459, respectively, using the PrimeSTAR Kit (Takara, Japan). The forward and reverse primers (F: 5′-CACCTCTAGAGAGGGCCTATTTCCCATGATTCCTTCATAT-3′, R: 5′-CACCGGTACCAAAAAAGCACCGACTCGGTGCCACTTTTTC-3′) were synthesized by Sangon Biotech (Shanghai). U6-26a-sg2-tracRNA and U6-26a-sg3-tracRNA sequences were inserted into 26a-sg1-PX459 by double endonucleases (Xba I and Kpn I) to construct a dual−sgRNA vector, named 26a-sg1-sg2-PX459 and 26a-sg1-sg3-PX459 (Figure 1C). 26b-sg1-sg2-PX459 and 26b-sg1-sg3-PX459 were obtained via the same method.

Zhu L., Jiao H., Gao W., Huang L., Shi C., Zhang F., Wu J, & Luo J. (2023). Fatty Acid Desaturation Is Suppressed in Mir-26a/b Knockout Goat Mammary Epithelial Cells by Upregulating INSIG1. International Journal of Molecular Sciences, 24(12), 10028.

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CHIKV E1 Gene Sequencing Protocol

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The ReverTra Ace kit (Toyobo, Osaka, Japan) was used for reverse transcription of the RNA samples that tested positive for CHIKV with real-time RT–PCR. Then, the E1 gene segment (294 bp) was amplified using the PrimeSTAR kit (Takara Bio, Shiga, Japan), and the amplicon DNA sequence was acquired by applying the Sanger method. The primers used [41 (link)] are documented in Supplementary Table S2. The nucleotide sequences were analysed using DNADynamo v. 1.63 (Blue Tractor Software). Then, the sequences were aligned with CHIKV global sequences using MAFFT v. 7.520 [44 (link)] and subjected to phylogenetic analysis using the maximum-likelihood method with 1000 bootstrap replicates in MEGA 11 [45 (link),46 (link)]. The Tamura–Nei and invariant site models were employed for this analysis after finding the best-fit model based on the Bayesian information criterion (BIC) using W–IQ–TREE [47 (link),48 (link),49 (link),50 (link)]. The nucleotide sequences obtained from current study were submitted to the GenBank database under accession numbers OR492236, OR492237, OR492238, OR492239, OR492240, OR492241, and OR492242.

Nguyen T.V., Ngwe Tun M.M., Cao M.T., Dao H.M., Luong C.Q., Huynh T.K., Nguyen T.T., Hoang T.N., Morita K., Le T.Q., Pham Q.D., Takamatsu Y, & Hasebe F. (2023). Serological and Molecular Epidemiology of Chikungunya Virus Infection in Vietnam, 2017–2019. Viruses, 15(10), 2065.

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RT-PCR for Quantitative HCV Detection

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The cDNA was synthesized from viral RNA in a 20 μL reaction volume including 5 μL RNA and 1 μL Random primer at 65 °C for 6 min, then 4 μL Reaction Buffer, 2 μL dNTP Mixture, 1 μL RNase inhibitor, 1 μL Reverse transcriptase, 6 μL RNase Free H₂O (Takara, Dalian, China) at 42 °C for 60 min, followed by 72 °C for 7 s. HCV fragments were amplified using a PrimeSTAR Kit (Takara, Dalian, China) in a 20 μL reaction volume including cDNA 2 μL, primer 1 μL, mix 10 μL, 7 μL ddH₂O was performed with 1 cycle 42 °C for 5 min, 95 °C 3 min, followed by 40 cycles, each consisting of 94 °C 30 s, 56 °C 50 s, 72 °C 1 min, then 72 °C 10 min, 4 °C 10 min. All manufacturer protocols were followed. Linear range is 1 × 10³–5 × 10⁷ IU/mL, The lowest sensitivity is 5 × 10² IU/mL.

Niu Z., Zhang P, & Tong Y. (2016). Age and gender distribution of Hepatitis C virus prevalence and genotypes of individuals of physical examination in WuHan, Central China. SpringerPlus, 5(1), 1557.

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CERK1 Mutagenesis and Silencing in Arabidopsis

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The Tyr-to-Phe site-directed mutagenesis of CERK1 was introduced by the overlap extension PCR with the PrimeSTAR Kit (Takara, Japan). The primers used are listed in Table S1. A 23FLAG coding sequence was inserted in-frame immediately preceding the stop codon of the CERK1 coding sequence. The CERK1 variants carrying individual point mutations were inserted into the pCAMBIA 1301 binary vector behind the CaMV 35S promoter or the native promoter of CERK1 (Kadota et al., 2014) . The CIPP-GFP chimeric gene was also cloned using the pCAMBIA 1301 binary vector containing the 35S promoter. The region of 1-477 bp within the coding sequence of CIPP1 was selected as the RNAi target to generate the CIPP1-RNAi silencing plants. This sequence was inserted into the pCAMBIA 1301 binary vector on both sides of an intron but in an opposite direction and its transcription was driven by the 35S promoter. Sequence-verified pCAMBIA binary plasmids were introduced into cells of Agrobacterium tumefaciens strain EHA105 by electroporation. Agrobacteria expressing CERK1 mutants were transformed into the cerk1 mutant plants, while those expressing the CIPP1-GFP or CIPP1-RNAi constructs were transformed into Col-0 by the floral dipping method (Zhang et al., 2006) . Positive transgenic plants were screened on 0.5 3 MS plates with 25 mg/L hygromycin and the T2 transgenic plants were used for further analyses.

Liu J., Liu B., Chen S., Gong B.Q., Chen L., Zhou Q., Xiong F., Wang M., Feng D., Li J.F., Wang H.B, & Wang J. (2018). A Tyrosine Phosphorylation Cycle Regulates Fungal Activation of a Plant Receptor Ser/Thr Kinase. Cell host & microbe, 23(2).

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