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Ssoadvanced sybr reagent

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The SsoAdvanced SYBR reagent is a real-time PCR reagent designed for sensitive and specific detection of target DNA sequences. It provides a simple and reliable solution for quantitative gene expression analysis.

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Lab products found in correlation

Qpcr platform, by Bio-Rad (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Cfx384 qpcr machine, by Bio-Rad (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)

4 protocols using ssoadvanced sybr reagent

1

Quantitative PCR Analysis of Gene Expression

Cited 1 time
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RNA was prepared from cultured tumor cells using RNeasy Mini Kit or RNeasy Micro Kit (Qiagen). cDNA was generated using High-capacity cDNA Reverse Transcription Kit from 1 μg RNA in 20 μl reaction volume and diluted 1:10 for qPCR analysis (Life Technologies). qPCR analysis was performed using 2 μl diluted cDNA with biological (2 (link)-3 (link)) and technical replicates (2 (link)-3 (link)) using SsoAdvanced SYBR reagent (Bio-Rad) and Bio-Rad qPCR platform, and results were normalized to the expression of Tbp using the Bio-Rad software. Primer sequences utilized for qPCR were: Usp22-F-CTC-CCC-ACA-CAT-TCC-ATA-CAA-G; Usp22-R-TGG-AGC-CCA-CCC-GTA-AAG-A; Atxn7l3-F-TTG-TCT-GGC-CTG-GAT-AAC-AGC; Atxn7l3-R-CCG-GTG-TAC-TTC-AAA-GCA-GAA-TC; Tbp-F-AGA-ACA-ATC-CAG-ACT-AGC-AGC-A; Tbp-R-GGG-AAC-TTC-ACA-TCA-CAG-CTC.
Li J., Yuan S., Norgard R.J., Yan F., Yamazoe T., Blanco A, & Stanger B.Z. (2019). Tumor cell–intrinsic USP22 suppresses antitumor immunity in pancreatic cancer. Cancer immunology research, 8(3), 282-291.
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2

Quantitative Real-Time PCR Optimization

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qPCR was performed at 10 μl/well using the Bio-Rad CFX 384 qPCR machine (Bio-rad). Each well contained: 3 μl diluted cDNA, 0.25 μl each of 10 μM forward and reverse primers (Supplementary Table 4), 1.5 μl H2O and 5 μl SsoAdvanced SYBR reagent (Bio-rad).
Katsuda T., Sussman J.H., Ito K., Katznelson A., Yuan S., Takenaka N., Li J., Merrell A.J., Cure H., Li Q., Rasool R.U., Asangani I.A., Zaret K.S, & Stanger B.Z. (2024). Cellular reprogramming in vivo initiated by SOX4 pioneer factor activity. Nature Communications, 15, 1761.
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3

Quantitative PCR Protocol for Gene Expression

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qPCR was performed at 10 μl/well using the Bio-Rad CFX 384 qPCR machine (Bio-rad). Each well contained: 3 μl diluted cDNA, 0.25 μl each of 10 μM forward and reverse primers (Table S6), 1.5 μl H2O and 5 μl SsoAdvanced SYBR reagent (Bio-rad).
Katsuda T., Sussman J., Ito K., Katznelson A., Yuan S., Li J., Merrell A.J., Takenaka N., Cure H., Li Q., Rasool R.U., Asangani I.A., Zaret K.S, & Stanger B.Z. (2023). Physiological reprogramming in vivo mediated by Sox4 pioneer factor activity. bioRxiv.
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4

Quantitative RT-PCR Analysis of Tumor Cell Genes

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RNA was prepared from cultured tumor cells or sorted cells from implanted tumors using RNeasy Mini Kit or RNeasy Micro Kit (Qiagen). cDNA was generated using High-capacity cDNA Reverse Transcription Kit (Life Technologies). qPCR analysis was performed using SsoAdvanced SYBR reagent (Bio-Rad) and Bio-Rad qPCR platform, and results were normalized to the expression of Tbp. Primer sequences utilized for qPCR were: Cxcl1: forward-CTG-GGA-TTC-ACC-TCA-AGA-ACA-TC, reverse-CAG-GGT-CAAGGC-AAG-CCT-C; Cxcl5:forward-TGC-CCT-ACG-GTG-GAA-GTC-ATA, reverse-TGC-ATT-CCG-CTTAGC-TTT-CTT-T; Csf-3: forward-ATG-GCT-CAA-CTT-TCT-GCC-CAG, reverse-CTG-ACA-GTG-ACCAGG-GGA-AC; Csf2: forward-GGC-CTT-GGA-AGC-ATG-TAG-AGG, reverse-GGA-GAA-CTC-GTTAGA-GAC-GAC-TT; Myc: forward-GCA-TGA-GGA-GAC-ACC-GCC-CA, reverse-GGT-TTG-CCT-CTT-CTC-CAC-AGA. As the qRT-PCR analyses of the tumor cells were performed in different runs, we used 6419c5 in every run as a sample for normalization across runs and compared across samples.
Li J., Byrne K.T., Yan F., Yamazoe T., Chen Z., Baslan T., Richman L.P., Lin J., Sun Y.H., Rech A.J., Balli D., Hay C.A., Sela Y., Merrell A.J., Liudahl S.M., Gordon N., Norgard R.J., Yuan S., Yu S., Chao T., Ye S., Eisinger-Mathason T.S., Faryabi R.B., Tobias J.W., Lowe S., Coussens L.M., Wherry E.J., Vonderheide R.H, & Stanger B.Z. (2018). Tumor cell-intrinsic factors underlie heterogeneity of immune cell infiltration and response to immunotherapy. Immunity, 49(1), 178-193.e7.
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