Rnx plus solution for total rna isolation

Manufactured by Sinaclon

Sourced in Iran, Islamic Republic of

RNX-Plus solution is a reagent designed for the isolation of total RNA from various biological samples. It utilizes a guanidinium thiocyanate-phenol-chloroform extraction method to effectively separate and purify RNA from the sample.

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Lab products found in correlation

Maxime rt premix kit, by iNtRON Biotechnology (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Pe annexin v 7 aad apoptosis detection kit, by BD (1 mentions) Sybr premix ex taq 2, by Thermo Fisher Scientific (1 mentions) Jc 1 mmp assay kit, by Cayman Chemical (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Merck Group (1 mentions) Lightcycler device, by Roche (1 mentions)

4 protocols using rnx plus solution for total rna isolation

RNA Extraction and cDNA Synthesis Protocol

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RNA extraction was done by RNX-Plus solution
for total RNA isolation (SinaClon Bioscience, Iran)
and quality control procedure of the isolated RNA was
undertaken with measurement of the absorbance at
260/280 nm by Biophotometer; the isolated RNA had
an optical density between 1.9-2 with double distilled
water used as blank. Afterward, cDNA was produced
by a GeneAll kit (HyperScript^TM Reverse Transcriptase,
South Korea); cDNA synthesis was done in 20 µl volume
containing 3 µl extracted RNA, 1 µl dNTP, 1 µl oligo dT
and 9 µl nuclease-free distilled water, which was heated
to 65°C for 5 minutes and then placed on ice. After
that, 6 µl of RT buffer including 10X RTase reaction
buffer, DTT, HyperScriptTM Reverse Transcriptase and
ZymAllTMRNase inhibitor were added and incubated for
50 minutes at 55°C followed by 5 minutes at 85°C.

Habibi H., Atashi A., Abroun S, & Noruzinia M. (2018). Synergistic Effect of Simvastatin and Romidepsin on Gamma-globin Gene Induction. Cell Journal (Yakhteh), 20(4), 576-583.

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RNA Extraction and RT-PCR Detection of SARS-CoV-2

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RNA from faeces was extracted using RNX‐Plus solution for total RNA isolation (Sinaclon Bioscience, Iran) as instructed by the manufacturer. Then the complementary DNA (cDNA) was produced by using the Maxime RT premix kit (Intron biotechnology, Korea) as instructed by the manufacturer.
The PCR conditions used were based on the two papers by of Hasoksuz, Kayar, Dodurka, and Ilgaz (2005) as well as Liu et al., (2006) for the gene S and the study of Fukuda et al., (2012) for the gene N. Briefly after initial denaturation at 94 C for 5 min, cDNAs entered to 40 cycles, each one with three steps including denaturation at 94 C for 45 s, primer annealing at 52 C (for N gene) or 50 C (for S gene) for 45 s, extension at 72 C for 1 min that finally followed by final extension step at 72 C for 10 min.
The PCR products were visualized on 1.5% agarose gel stained with ethidium bromide. PCR the products 407 bp and 654 bp were detected for genes N and S, respectively.

Lotfollahzadeh S., Madadgar O., Reza Mohebbi M., Reza Mokhber Dezfouli M, & George Watson D. (2020). Bovine coronavirus in neonatal calf diarrhoea in Iran. Veterinary Medicine and Science, 6(4), 686-694.

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Cytotoxicity and Apoptosis Analysis

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Roswell Park Memorial Institute medium (RPMI-1640) and fetal calf serum (FCS) were purchased from Gibco (Ashland, KY). Cell culture grade dimethyl sulfoxide (DMSO), 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), trypan blue dye and propidium iodide (PI) were purchased from Sigma (St. Louis, MO). PE Annexin V/7-AAD apoptosis detection kit was obtained from BD Pharmingen™ (San Diego, CA) and JC-1 MMP assay kit from Cayman Chemical (Ann Arbor, MI). RNX-Plus solution for total RNA isolation was obtained from Sinaclon (Tehran, Iran). SYBR Premix Ex Taq II and high-capacity cDNA reverse transcription kit were provided by Applied Biosystems (ABI, Foster City, CA).

Asemani Y., Azadmehr A., Hajiaghaee R, & Amirghofran Z. (2018). Anticancer potential of Ferula hezarlalehzarica Y. Ajani fraction in Raji lymphoma cell line: induction of apoptosis, cell cycle arrest, and changes in mitochondrial membrane potential. DARU Journal of Pharmaceutical Sciences, 26(2), 143-154.

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Quantitative Assessment of GFAP, NSE, and RAGE mRNA

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The method was used to assess the expression of GFAP, NSE, and RAGEs mRNA. Brain tissue was isolated and stored at −70°C for RNA extraction. RNX‐Plus solution for total RNA isolation (Sinaclon, Iran, Cat. No.: EX6101) was used to extract of RNA from each sample. The concentration of the extracted RNA was determined using NanoDrop1 C instrument (Thermo Scientific Co., USA). The amount of RNA in the samples was normalized with DEPC water. The Easy™ cDNA Synthesis Kit (Para Tus Inc., Iran, Cat. No.: A101161) was used for cDNA synthesis. Genes expression in each sample was quantified by RT‐qPCR. 16S rRNA was used as the reference internal control gene. The RT‐qPCR mixture contained 12.5 mL SYBR® green real‐time PCR master mix (Para Tus Inc., Iran, Cat. No.: C101022), 0.8 mL cDNA, 9.7 mL DNase‐free water, and 1 mL for each forward and reverse primer. The RT‐qPCR reaction was performed using a Light Cycler device (Roche, Germany). The RT‐qPCR conditions were as follows: 94°C for 5 min as initial denaturation followed by 40 cycles of 95°C for 30 s, 56°C for 30 s, and 72°C for 30 s.

Basatinya A.M., Sajedianfard J., Nazifi S, & Hosseinzadeh S. (2024). The analgesic effects of insulin and its disorders in streptozotocin‐induced short‐term diabetes. Physiological Reports, 12(8), e16009.

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