Miseq 300 bp paired reads platform

Manufactured by Illumina

Sourced in United States

The MiSeq 300-bp paired-reads platform is a benchtop sequencing system designed for targeted sequencing applications. It generates up to 15 Gb of sequence data per run with read lengths of up to 300 base pairs in a paired-end configuration.

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Lab products found in correlation

Lysing matrix b tube, by MP Biomedicals (2 mentions) Axyprep dna gel extraction kit, by Corning (1 mentions) Fastdna spin kit, by MP Biomedicals (1 mentions) Nextflex rapid dna seq kit, by PSC Biotech (1 mentions) Nanodrop system, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MiSeq platform (7007 citations) MiSeq 300 (25 citations) MiSeq 300 bp (11 citations) MiSeq reads (3 citations) Miseq (300-bp paired-end reads) platform (2 citations)

3 protocols using miseq 300 bp paired reads platform

Microbial Diversity Profiling by 16S rRNA Sequencing

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DNA was extracted from the skin microbial samples using the commercial FastDNA Spin Kit (MP Biomedicals, USA) following the instructions provided by the manufacturer. The V3-V4 variable region in the 16S rRNA gene was PCR-amplified with the primers 338F (5′-ACTCCTACGGGAGGCAGCAG-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) using a thermocycler (GeneAmp 9700, ABI). PCR amplicons were purified using the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences, USA), libraries were prepared using the NEXTFLEX Rapid DNA-Seq Kit (Bioo Scientific, USA), and sequencing was performed using the Illumina MiSeq 300-bp paired-reads platform (Illumina, USA) in accordance with standard protocols provided by Majorbio Bio-Pharm Technology Co. Ltd. (Shanghai, China) [29 ].

Ma L., Jiang H., Han T., Shi Y., Wang M., Jiang S., Yang S., Yao L., Jia Q, & Shao L. (2023). The menstrual cycle regularity and skin: irregular menstrual cycle affects skin physiological properties and skin bacterial microbiome in urban Chinese women. BMC Women's Health, 23, 292.

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Metagenomic DNA Extraction and 16S Sequencing

Cited 2 times

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Metagenomic DNA was extracted from skin swab samples as previously published [15 (link), 16 (link)]. Samples were thawed and transferred aseptically to Lysing Matrix B tubes (MP Biomedicals, Solon, OH, USA). Enzymatic bacterial lysis was conducted using lysozyme, mutanolysin, proteinase K, and lysostaphin, followed by mechanical lysis through bead beating. A Zymo fecal DNA kit (Zymogen, Irvine, CA, USA) was used to further purify metagenomic DNA. DNA quality assurance was performed with spectrophotometric measurements on the NanoDrop system and gel electrophoresis. Negative extraction phosphate-buffered saline controls were also included in sample processing to confirm that contaminant DNA was not introduced into samples during the extraction process. Following extraction, the V3V4 hypervariable region of the 16S rRNA gene was polymerase chain reaction amplified and sequenced on an Illumina MiSeq 300-bp paired-reads platform (Illumina, San Diego, CA, USA) as previously described [17 (link)].

Rainer B.M., Thompson K.G., Antonescu C., Florea L., Mongodin E.F., Bui J., Fischer A.H., Pasieka H.B., Garza L.A., Kang S, & Chien A.L. (2020). Characterization and Analysis of the Skin Microbiota in Rosacea: A Case–Control Study. American journal of clinical dermatology, 21(1), 139-147.

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Metagenomic DNA Extraction from Skin Swabs

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Extraction of metagenomic DNA (mgDNA) was conducted from skin swabs as previously described20 (link)21 (link). Thawed samples were transferred aseptically to Lysing Matrix B tubes (MP Biomedicals, Solon, OH, USA). Lysozyme, mutanolysin, proteinase K, and lysostaphin enzymatic bacterial lysis was followed by bead-beating mechanical lysis. mgDNA was further purified using a Zymo fecal DNA kit (Zymogen, Irvine, CA, USA). Spectrophotometric measurements on the NanoDrop system (NanoDrop Technologies, Wilmington, DE, USA) and gel electrophoresis were used for DNA quality assurance. Phosphate-buffered saline negative extraction controls were included in sample processing. The V3V4 hypervariable region of the 16S rRNA gene was PCR amplified. The Illumina MiSeq 300-bp paired-reads platform (Illumina, San Diego, CA, USA) was used for sequencing as previously published22 (link).

Thompson K.G., Rainer B.M., Antonescu C., Florea L., Mongodin E.F., Kang S, & Chien A.L. (2020). Minocycline and Its Impact on Microbial Dysbiosis in the Skin and Gastrointestinal Tract of Acne Patients. Annals of Dermatology, 32(1), 21-30.

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