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Allstars neg control sirna

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The AllStars Neg Control siRNA is a laboratory reagent designed to serve as a negative control in RNA interference (RNAi) experiments. It is a synthetic, non-targeting small interfering RNA (siRNA) that does not recognize any known mammalian gene.

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Lab products found in correlation

Flexitube sirna, by Qiagen (2 mentions) Allstars hs cell death control sirna, by Qiagen (1 mentions) Rnaimax, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Hyperfect transfection reagent, by Qiagen (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)

4 protocols using allstars neg control sirna

1

Silencing YBEY Gene in Breast Cancer Cells

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MDA-MB-231, T47D, and MCF-7 cells were plated at 1.25 × 105 cells/well in 6-well plates and reverse-transfected using three siRNAs (Y1, Y2, and Y3) targeting different sites of YBEY mRNA (Dharmacon, Lafayette, CO, USA) at 10 nM using liposomal delivery (RNAiMAX, Life Technologies, Carlsbad, CA, USA). The sequences for each siRNA have been provided in Table S2. A non-targeting control (NTC) siRNA (AllStars Neg Control siRNA, Qiagen, Germantown, MD, USA) and a positive control (POS) siRNA (AllStars Hs Cell Death Control siRNA, Qiagen) were used as the negative control and the positive control, respectively. The k.d. efficiency was assessed after 24 h and up to 48 h post-transfection by quantitative real-time PCR (qPCR) using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a housekeeping gene. These data represent fold-changes normalized to the negative control and have been provided in Figure S2.
Shidal C., Shu X., Wu J., Wang J., Huang S., Long J., Bauer J.A., Ping J., Guo X., Zheng W., Shu X.O, & Cai Q. (2021). Functional Genomic Analyses of the 21q22.3 Locus Identifying Functional Variants and Candidate Gene YBEY for Breast Cancer Risk. Cancers, 13(9), 2037.
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2

Silencing Integrin α4 in Melanoma

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The siRNA directed against the human integrin subunit α4 (Hs_ITGA4_6; Qiagen, Hilden, Germany) as well as the non-targeting control siRNA (AllStars Neg. Control siRNA; Qiagen) were diluted according to the manufacturer’s instructions and stored at -20°C. Melanoma cells were transfected with siRNA at a concentration of 125 nM using the Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer’s instructions. Cells were added to the siRNA–lipofectamine suspension and DMEM without antibiotics. The transfection was repeated after 24 h of incubation. Transfected cells were harvested after a further incubation of 72 h and directly used in experiments with parallel flow cytometry analysis of α4-integrin expression.
Amschler K., Koßmann E., Erpenbeck L., Kruss S., Schill T., Schön M., Möckel S.M., Spatz J.P, & Schön M.P. (2017). Nanoscale Tuning of VCAM-1 Determines VLA-4-dependent Melanoma Cell Plasticity on RGD Motifs. Molecular cancer research : MCR, 16(3), 528-542.
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3

siRNA-Mediated Knockdown of Il6 and Pdcd1 in H9c2 Cells

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Small interfering RNA (siRNA)-Il6 (siIl6) and siRNA-Pdcd1 (siPdcd1) were transfected into H9c2 cells using the HyperFect transfection reagent (Qiagen) according to the manufacturer's protocol. A non-targeting siRNA (Mock) was used as a vehicle control to assess the non-sequence-specific effects of transfected siRNAs. The siRNAs used were siIl6, a FlexiTube siRNA (ID no. SI01525356, Qiagen) and siPdcd1, a Silencer® Select Pre-designed siRNA Product (ID no. s154115, Thermo Fisher Scientific, Inc.); the negative control siRNA was obtained as AllStars Neg. Control siRNA (ID no. AM4611, Qiagen). Transfections consisted of 4×104 cells combined with a given siRNA (final concentration, 10 nM) in the presence of HyperFect reagent; the mixtures then were incubated for 24 h before assessment of Il6 or Pdcd1 expression.
Kanno S.I, & Hara A. (2020). The mRNA expression of Il6 and Pdcd1 are predictive and protective factors for doxorubicin-induced cardiotoxicity. Molecular Medicine Reports, 23(2), 113.
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4

ROCK1 Knockdown in BPXV Replication

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In order to further confirm the role of ROCK1 in BPXV replication, as well as to avoid the possibilities that ROCK1 inhibitor may have some off-target effects in the host cells, ROCK1 was knockdown from the cells in a sequence dependent manner by using small interfering RNA (siRNA). Briefly, Vero cells were grown in 96 well plates. When the cells were at ∼75% confluency, 50 nmol and 100 nmol of ROCK1 (FlexiTube siRNA, Qiagen, Germany] or control siRNA (AllStars Neg Control siRNA, Qiagen, Germany] were transfected using Lipofectamine 3000 as per the instruction of manufacturer (Invitrogen, Carlsbad, USA). At 48 h post-transfection, cells were infected with BPXV at MOI of 1 and virus released in the infected cell culture supernatant at 48 hpi was quantified by plaque assay.
Kumar R., Chander Y., Khandelwal N., Verma A., Rawat K.D., Shringi B.N., Pal Y., Tripathi B.N., Barua S, & Kumar N. (2022). ROCK1/MLC2 inhibition induces decay of viral mRNA in BPXV infected cells. Scientific Reports, 12, 17811.
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