3730xl system

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States

The 3730XL system is a DNA sequencing platform designed for high-throughput DNA analysis. It utilizes capillary electrophoresis technology to accurately and efficiently determine the nucleotide sequence of DNA samples. The system is capable of processing multiple samples simultaneously, making it a versatile tool for various applications in the field of genomics and molecular biology.

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Lab products found in correlation

Methylamp dna modification kit, by Epigentek (1 mentions) Dneasy blood tissue kit, by Qiagen (1 mentions) Ez dna methylation direct kit, by Zymo Research (1 mentions) Ex taq dna polymerase, by Bio-Rad (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Genejet plasmid miniprep kit, by Thermo Fisher Scientific (1 mentions) Pgl3 basic firefly luciferase vector, by Promega (1 mentions) Prl sv40 renilla vector, by Promega (1 mentions) Dual luciferase reporter assay system kit, by Promega (1 mentions) X tremegene hp dna transfection reagent, by Roche (1 mentions)

3 protocols using 3730xl system

Methylation Analysis of KLF4 in OSCC

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Genomic DNA was extracted from OSCC cell lines using the DNeasy Blood & Tissue Kit (Qiagen, Valencia, CA), according to the manufacturer's instructions. Genomic DNA was bisulfite-modified with the EZ DNA Methylation-Direct Kit (Zymo Research Co., Orange, CA), and the modified DNA was amplified and examined by electrophoresis in a 1.5% agarose gel to confirm that a single band had been obtained. It was then sequenced with the 3730 xl system (Applied Biosystems, Warrington, United Kingdom). The bisulfite sequencing results were analyzed with BISMA, an online software tool.
The methylation status of KLF4 was further validated in healthy oral mucosa, hyperplasia, dysplasia and OSCC samples. Bisulfite conversion was carried out using the Methylamp DNA Modification Kit (Epigentek, Farmingdale, NY). Methylation analysis was performed using a fluorescence-based, real-time PCR assay with the following conditions: 95°C for 10 min, 35 cycles (95°C 15 s, 60°C 60 s), then 72°C for 7 min.

Li W., Liu M., Su Y., Zhou X., Liu Y, & Zhang X. (2015). The Janus-faced roles of Krüppel-like factor 4 in oral squamous cell carcinoma cells. Oncotarget, 6(42), 44480-44494.

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DNAH6 Gene Mutation Detection

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Forward and reverse PCR primers targeting mutation regions in the DNAH6 gene had the following sequences: c.6582 C > A: 5′-CCATCTTGTGCCCTGACAGT-3′ and 5′-GTCTGTCCACCCTCTGAAGC-3′; c.11258 G > A: 5′-GGAAGGAAATTCTGTTGTGTGCT-3′ and 5′-TGGGGATAGGGTGTGGCTATAA-3′; c.10025 G > A: 5′-CTGAATGCGAACAAGGGAGAC-3′ and 5′-AGATAGGGTTCCTTGCTGACG-3′; and c.2823dupT: 5′-TGAAAGGAATAAACAGGTGATGCT-3′ and 5′-ACTCTGAATGCCCCTCCTAAC-3′. These regions were amplified by PCR using Ex Taq DNA polymerase (Bio-Rad, Hercules, CA, USA) in all patients and their family members. PCR products were sequenced on a 3730XL system (Applied Biosystems, Foster City, CA, USA) according to the manufacturer’s instructions.

Tu C., Nie H., Meng L., Yuan S., He W., Luo A., Li H., Li W., Du J., Lu G., Lin G, & Tan Y.Q. (2019). Identification of DNAH6 mutations in infertile men with multiple morphological abnormalities of the sperm flagella. Scientific Reports, 9, 15864.

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Regulation of PD-L1 Promoter Activity

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The reporter gene constructs for PD‐L1 (−373/+328) were generated by PCR amplification. The PCR products were purified with a high PCR product purification kit (GeneJET Plasmid Miniprep kit; Thermo Fisher Scientific) and cloned into the pGL3‐basic firefly luciferase vector (Promega). The sequence of each cloned promoter region was confirmed by sequencing (Applied Biosystems 3730XL system). For reporter assays, cells were cotransfected with the firefly luciferase construct pRL‐SV40 Renilla vector (Promega) and the PD‐L1 promoter construct vector using X‐tremeGENE HP DNA Transfection Reagent (6366236001; Roche). After 6 h, cells were treated with 10 ng/ml TGF‐β (PHG9214; Fisher Scientific), 10 ng/ml IL‐1β (PHC0811; Fisher Scientific), 10 ng/ml IL‐6 (PHC0061; Fisher Scientific), 10 ng/ml EGF (PHG0311, Fisher Scientific), 10 ng/ml IFN‐α (PHC4014, Fisher Scientific), 10 ng/ml IFN‐β (PHC4244, Fisher Scientific), 10 ng/ml IFN‐γ (PHC4031, Fisher Scientific), or 10 ng/ml TNF‐α (PHC3015, Fisher Scientific). Then after 24 h, luciferase activity was measured by using the Dual‐Luciferase Reporter Assay System kit (Promega). A549‐hACE2 cells were transfected with PD‐L1 promoter plasmid expressing luciferase. After that, A549‐hACE2 cells were incubated with SARS‐CoV‐2 wild‐type, and Omicron variants spike‐pseudotyped lentivirus for 48 h followed by reporter assays.

Huang H., Wang S., Fang G., Chou W., Liao C., Sun C., Jan J., Ma H., Ko H., Ko Y., Chiang M., Liang J., Kuo C., Lee T., Morales‐Scheihing D., Shen C., Chen S., McCullough L.D., Cui L., Wernig G., Tao M., Lin Y., Chang Y., Wang S., Lai Y, & Li C. (2023). Upregulation of PD‐L1 by SARS‐CoV‐2 promotes immune evasion. Journal of Medical Virology, 95(2), e28478.

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