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Exgene cell sv kit

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The Exgene Cell SV Kit is a DNA extraction kit designed for the rapid and efficient purification of genomic DNA from various cell types. The kit utilizes a simple and straightforward procedure to isolate high-quality DNA suitable for downstream applications such as PCR, sequencing, and analysis.

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Lab products found in correlation

Emeraldamp pcr master mix, by Takara Bio (2 mentions) Plasmid sv kit, by GeneAll (2 mentions) Oligonucleotides, by Cosmogenetech (2 mentions) T4 polynucleotide kinase, by Takara Bio (2 mentions) T4 dna ligase, by New England Biolabs (2 mentions) Q5 high fidelity dna polymerase, by New England Biolabs (2 mentions) Restriction endonuclease, by New England Biolabs (2 mentions) Luria bertani media, by BD (2 mentions) Geneamp pcr system, by Thermo Fisher Scientific (2 mentions) Pcr purification kit, by Thermo Fisher Scientific (2 mentions) Gel logic 200 imaging system, by Kodak (2 mentions) Q5 polymerase, by New England Biolabs (2 mentions) Exprep plasmid sv kit, by GeneAll (2 mentions) Qubit fluorometer, by Thermo Fisher Scientific (1 mentions) T gradient thermal cycler, by Analytik Jena (1 mentions) Depc dw, by Bioneer (1 mentions) Premix ex taq, by Takara Bio (1 mentions) Novaseq 6000 system, by Illumina (1 mentions) Pcr clean up system, by Promega (1 mentions) Veriti 96 well thermal cycler, by Thermo Fisher Scientific (1 mentions)

28 protocols using exgene cell sv kit

1

Bacterial Genomic DNA Extraction and PCR Analysis

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The genomic DNA was extracted from the bacterial cells using the GeneAll ExgeneTM Cell SV kit (GeneAll Biotechnology, Seoul, Korea) according to the manufacturer’s protocol. DNA concentrations were determined with a QubitTM fluorometer (Invitrogen®, Carlsbad, CA, USA).
The PCR assays used 1 μl of template DNA in a 25 μl reaction mixture containing 2 mM of Tris-HCl (pH 8.0), 10 mM of KCl, 10 uM of EDTA, 100 uM of DTT, 0.05% Tween 20, 0.05% Nonidet P-40, 5% glycerol, 2.5 mM of dNTP, 5 units of Ex Taq DNA (TaKaRa Bio Inc., Shiga, Japan), and 10 pM of each primer. The reactions were performed in a T Gradient Thermal Cycler (Biometra, Göttingen, Germany) programmed for one cycle of 10 min at 95°C, followed by 35 cycles of 30 s at 95°C, 30 s at 60°C, and 30 s at 72°C, with a final extension step for 7 min at 72°C. The amplicons were analyzed by electrophoresis in a 2% agarose gel in TAE buffer.
The TaqMan PCR assay was carried out using 2 μl of DNA template, 8.5 μl of Premix Ex Taq (TaKaRa Bio Inc., Shiga, Japan), 1 μl of TaqMan probe (10 pmoles/μl), 0.5 μl (10 pmoles/μl) of each nested primer, and 12.5 μl of DEPC-DW (Bioneer, Daejeon, Korea). Real-time PCR amplifications were performed in a Smart CyclerTM System (Cepheid, Sunnyvale, CA, USA) with 40 cycles at 95°C for 30 sec and 60°C for 20 sec after initial incubation for 30 sec at 95°C.
An J.H., Noh Y.H., Kim Y.E., Lee H.I, & Cha J.S. (2015). Development of PCR and TaqMan PCR Assays to Detect Pseudomonas coronafaciens, a Causal Agent of Halo Blight of Oats. The Plant Pathology Journal, 31(1), 25-32.
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2

Time-course Real-time PCR for HHVs

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Overall, 1, 3, 7 and 14 days post-infection, the cells were harvested and DNA was extracted with the GeneAll ExgeneTM Cell SV kit (GeneAll Biotechnology, Seoul, Korea). Real-time PCR for HHVs was performed (as reported in Supplementary Table S2) following the manufacturer’s procedures.
Bortolotti D., Gentili V., Bortoluzzi A., Govoni M., Schiuma G., Beltrami S., Rizzo S., Baldi E., Caselli E., Pugliatti M., Castellazzi M., Fernández M., Fainardi E, & Rizzo R. (2022). Herpesvirus Infections in KIR2DL2-Positive Multiple Sclerosis Patients: Mechanisms Triggering Autoimmunity. Microorganisms, 10(3), 494.
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3

Bacterial Genome Sequencing Workflow

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The bacterial chromosomal DNAs from EPWSV and EPWSV2 were extracted from an overnight cultured sample using GeneAll ExgeneTM Cell SV kit (GeneAll Biotechnology) according to the manufacturer’s protocol. Sequencing libraries were constructed using the TruSeq DNA Nano DNA High Throughput Library Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer’s instructions. Samples were sequenced using the Illumina NovaSeq 6000 system (DNA Link, Seoul, Korea). Sequencing reads were then mapped onto the reference genome (GCF_000010245.2) using Burrows-Wheeler Aligner (version 0.7.12.) [48 (link)].
Gwon D.A., Seok J.Y., Jung G.Y, & Lee J.W. (2021). Biosensor-Assisted Adaptive Laboratory Evolution for Violacein Production. International Journal of Molecular Sciences, 22(12), 6594.
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4

Extraction of Genomic DNA from Leishmaniasis Samples

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Totally, 15 clinical samples sensitive to glucantime and 15 clinical samples resistant to glucantime were collected from the skin biopsies of the ACL patients, in the form of stained smears from different geographical locations in Iran such as Mashhad, Kerman, Esfahan and Ilam in 2017. The stain of the smear was removed in alcohol 96%; then, they were collected from the slides using the distilled water. Then, 200 μl of each sample were put in a 1.5 ml tube. Finally, the DNA genetic material was extracted using the GeneAll® Exgene TMCell SV kit (GeneAll BiotechnologyCo. Ltd., Seoul, South Korea).
FOZONGARI F., DALIMI A., ARAB S.S., BEHMANESH M, & KHAMMARI A. (2020). Trypanothione Reductase Gene Mutations in Meglumine Antimoniate Resistant Isolates from Cutaneous Leishmaniasis Patients Using Molecular Dynamics Method. Iranian Journal of Parasitology, 15(4), 511-520.
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5

Exon 3 SIRPA Amplification and Sequencing

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Genomic DNA was extracted from healthy donor PBMCs using a GeneAll Exgene Cell SV kit (GeneAll Biotechnology, Seoul, Korea). Exon 3 of SIRPA was amplified using EconoTaq PLUS GREEN 2X master mix (Lucigen Corporation, Middleton, WI, USA). The PCR thermal cycle was conducted on a Veriti 96-well thermal cycler (Applied Biosystems, Waltham, MA, USA) as follows: initial denaturation at 94 °C for 2 min; 35 cycles of 94 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min 40 s; final extension at 72 °C for 7 min. Amplicons were purified using Wizard SV gel and a PCR clean-up system (Promega), and the sequences were confirmed by Sanger sequencing. The primers used for PCR amplification and Sanger sequencing are as follows: PCR, (forward) 5′-AAACACACTGGCACGAGTCTA-3′, (reverse) 5′-GCCTTAACGGAGGAACCCAA-3′; Sanger sequencing, (forward) 5′-AGAATACAGGCTCATGTTGCAGGT-3′, (reverse) 5′-GCCTTCAGCAAATAGCATGACGT-3′.
Park H.R., Kim S.E., Keam B., Chung H., Seok S.H., Kim S., Kim M., Kim T.M., Doh J., Kim D.W, & Heo D.S. (2022). Blockade of CD47 enhances the antitumor effect of macrophages in renal cell carcinoma through trogocytosis. Scientific Reports, 12, 12546.
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6

Genomic DNA Extraction and Mutation Analysis

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Genomic DNA was extracted from cells grown in LB medium using a GeneAll Exgene™ Cell SV kit (GeneAll Biotechnology, Seoul, Korea). Pair-end libraries were prepared using the KAPA HyperPlus Kit (KAPA Biosystems, Wilmington, MA, USA). Raw reads were obtained using a MiniSeq 300-cycle Mid-Output kit on the MiniSeq system (Illumina, San Diego, CA, USA). Mutations were identified using the Breseq analysis software (version 0.33.2) [70 (link)]. A new reference genome of the VXA0 strain was generated based on Vibrio sp. dhg (NCBI accession number: CP028943.1, CP028944.1, and CP028945.1) to include the introduced xylA and to identify mutations in this strain. All genomes were sequenced with at least 25 × sequencing coverage. Raw sequencing files were deposited at SRA (Bioproject number: PRJNA720008). All discovered mutations were validated by Sanger sequencing.
Woo S., Lim H.G., Han Y.H., Park S., Noh M.H., Baek D., Moon J.H., Seo S.W, & Jung G.Y. (2022). A Vibrio-based microbial platform for accelerated lignocellulosic sugar conversion. Biotechnology for Biofuels and Bioproducts, 15, 58.
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7

DNA Purification and Molecular Cloning

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Plasmid and genomic DNA were purified using GeneAll® Plasmid SV kit and GeneAll® Exgene™ Cell SV kit (GeneAll Biotechnology, Seoul, Korea), respectively. Q5® High-Fidelity DNA Polymerase, restriction endonucleases, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA, USA). T4 Polynucleotide Kinase and EmeraldAmp® PCR Master Mix were purchased from Takara Bio Inc. (Shiga, Japan). Oligonucleotides were synthesized by Cosmogenetech (Seoul, Korea) (Additional file 1: Table S3). Other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless specified.
Kim D.H., Hwang H.G, & Jung G.Y. (2022). Optimum flux rerouting for efficient production of naringenin from acetate in engineered Escherichia coli. Biotechnology for Biofuels and Bioproducts, 15, 90.
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8

Genomic DNA Extraction from E. coli and NA12878

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We cultured E. coli EcNR2 and EcHB3 cells in Luria-Bertani (LB) media (BD Biosciences, USA) at 30°C in a shaking incubator. We harvested the cells by centrifugation and precipitated the genomic DNA using the GeneAll Exgene Cell SV Kit (GeneAll, Korea) according to the manufacturer's protocol. We purchased genomic DNA of NA12878 from the Coriell Institute (USA).
Lee J., Lim H., Jang H., Hwang B., Lee J.H., Cho J., Lee J.H, & Bang D. (2018). CRISPR-Cap: multiplexed double-stranded DNA enrichment based on the CRISPR system. Nucleic Acids Research, 47(1), e1.
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9

Plasmid and Genomic DNA Purification

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Plasmid and genomic DNA were purified using GeneAll® Plasmid SV kit and GeneAll® Exgene™ Cell SV kit (GeneAll Biotechnology, Seoul, Korea), respectively. Q5® High-Fidelity DNA Polymerase, restriction endonucleases, and T4 DNA ligase were purchased from New England Biolabs (Ipswich, MA, USA). T4 Polynucleotide Kinase and EmeraldAmp® PCR Master Mix were purchased from Takara Bio Inc. (Shiga, Japan). Oligonucleotides were synthesized by Cosmogenetech (Seoul, Korea). Other reagents were purchased from Sigma–Aldrich (St. Louis, MO, USA) unless specified.
Kim D.H., Hwang H.G., Ye D.Y, & Jung G.Y. (2024). Transcriptional and translational flux optimization at the key regulatory node for enhanced production of naringenin using acetate in engineered Escherichia coli. Journal of Industrial Microbiology & Biotechnology, 51, kuae006.
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10

Bacterial 16S rRNA Gene Amplification

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Bacterial genome was extracted from 1 mL 18 hrs old culture using GeneAll Exgene Cell SV kit (GeneAll Biotechnology Co. Ltd., Seoul, Korea), and the 16S rRNA gene was amplified by PCR. The universal primers 27F (5′-AGAGTTTGATCMTGGCTCAG-3′) and 1492R (5′-TACGGYTACCTTGTTACGACTT-3′) were used for the PCR amplification of the 16S rRNA gene. Then, PCR products were purified using HiYield Gel/PCR DNA Mini kit (Real Biotech Corporation, Minsheng Rd, Banqiao City, Taiwan) after separating them electrophoreticaly at 50 V on 0.8% (w/v) agarose gel. Molecular sizes of the PCR products were estimated by comparison with 1 kb DNA marker (BioNeer, Munpyeongseo Rd., Daejeon, Korea). Nucleotide sequences were determined at Macrogene Korea (Beotkkot Rd, Seoul, Korea) through cycle extension in an ABI 373 DNA sequencer (Applied Biosystems, Foster City, CA, USA). The sequencing results of each primer were imported to the BioEdit software program for assembly and alignment to generate reliable contigs after editing and removing ambiguous sequences. Finally, sequence similarities of each contig were examined by searching their homologies in the GeneBank database using BLAST. The hits after analyses were used to identify each isolate at a species level.
Gebru Y.A, & Sbhatu D.B. (2020). Isolation and Characterization of Probiotic LAB from Kimchi and Spontaneously Fermented Teff (Eragrostis tef (Zucc.) Trotter) Batter: Their Effects on Phenolic Content of Teff during Fermentation. BioMed Research International, 2020, 4014969.
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