Log In Sign up
The largest database of trusted experimental protocols

Rnaiso plus reagent kit

Manufactured by Takara Bio
Visit Supplier
Sourced in China, Japan

RNAiso Plus is a reagent kit designed for the isolation of total RNA from a variety of biological samples. The kit utilizes a guanidinium thiocyanate-phenol-chloroform extraction method to effectively lyse cells and denature proteins, allowing for the purification of high-quality RNA.

Automatically generated - may contain errors

Lab products found in correlation

Primescript rt reagent kit, by Takara Bio (5 mentions) Sybr premix ex taq kit, by Takara Bio (3 mentions) Sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions) Tb green advantage qpcr premix, by Takara Bio (2 mentions) Tb green premix ex taq tli rnaseh plus, by Takara Bio (2 mentions) Transscript 2 one step gdna removal and cdna synthesis supermix kit, by Transgene (2 mentions) Mir x mirna first strand synthesis kit, by Takara Bio (2 mentions) Sybr premix ex taq kit, by Thermo Fisher Scientific (2 mentions) Abi prism 7500 real time pcr system, by Thermo Fisher Scientific (2 mentions) Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions) Dnase kit, by Takara Bio (1 mentions) Tubulin, by Cell Signaling Technology (1 mentions) Baicalin, by Merck Group (1 mentions) Anti c caspase3, by Cell Signaling Technology (1 mentions) Anti sp1, by Cell Signaling Technology (1 mentions) Anti c parp, by Cell Signaling Technology (1 mentions) Mithramycin a, by Merck Group (1 mentions) Annexin 5 fitc, by Beyotime (1 mentions) Cck 8 kit, by Beyotime (1 mentions) Primescript reverse transcription reagent kit, by Takara Bio (1 mentions)

19 protocols using rnaiso plus reagent kit

1

Quantitative Analysis of SESA and EM1 Transcripts

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Samples were collected, frozen in liquid nitrogen, and stored at −79°C until use. For RNA isolation, the collected samples were ground to fine powder in liquid nitrogen. Total RNA was isolated using a RNAiso Plus Reagent Kit (Takara, Japan) and further purified with a DNase Kit (Takara, Japan). RT-qPCR was performed according to our previous study (Li et al., 2019 (link)). The primers for detecting SESA3 mRNA were 5’-AAGACAGAGATGCCAGAA-3’ and 5’-CCTCATTTGCTTGCTCAT-3.’ The primer set for detecting SESA4 mRNA was 5’-ACCAGAGCAAGTCAGGAA-3’ and 5’-TTAGTAGTAAGAAGGGATTGAAGG-3.’ The primer set for detecting EM1 mRNA was 5’-GGACTCAGTACGATGGAA-3’ and 5’-TTGTTGGTGAACTTTGACT-3.’ The primers for detecting the MDN1 mRNA were reported in our previous study (Li et al., 2016 (link)).
Li P.C., Ma J.J., Zhou X.M., Li G.H., Zhao C.Z., Xia H., Fan S.J, & Wang X.J. (2019). Arabidopsis MDN1 Is Involved in the Establishment of a Normal Seed Proteome and Seed Germination. Frontiers in Plant Science, 10, 1118.
+ Open protocol
+ Expand
2

Baicalin and Mithramycin-A Apoptosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Baicalin and mithramycin-A were procured from Sigma-Aldrich (St Louis, Missouri, USA). The annexin V-FITC and CCK-8 kits for detecting apoptosis were obtained from Beyotime (Shanghai, China), the RNAiso Plus reagent kit and the PrimeScript RT reagent kit (Perfect Real Time) were obtained from TAKARA, BIO (Kusatsu, Japan), whereas anti-sp1, anti-C-PARP, anti-C-caspase-3, and tubulin antibodies were obtained from Cell Signaling Technology (Danvers, Massachusetts, USA).
Ma W., Liu X, & Du W. (2019). Baicalin induces apoptosis in SW480 cells through downregulation of the SP1 transcription factor. Anti-Cancer Drugs, 30(2), 153-158.
+ Open protocol
+ Expand
3

Quantitative Gene Expression Analysis of U87 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
As directed by the manufacturer, RNA was extracted from U87 cells using the RNA isoPlus® Reagent Kit (Takara Biotechnology, Japan). To convert RNA into cDNA, the PrimeScript® RT Reagent Pack (Takara, Shiga, Japan) was used. Using the SYBR® Premix Ex Taq™ Unit and the 7500 Ongoing PCR Framework (Applied Biosystems, 7500 Continuous PCR Framework, Thermo, USA), the cDNA was enhanced. The conditions for cycling were as follows: The results were analyzed using the comparative Ct technique, with GAPDH acting as the loading control for the target genes during the forty cycles of 30 s at 95°C and 34 s at 60°C. The preliminaries were as per the following: IQGAP1 (Forward: 5′-ACCGTGGACCCAAAGAAC-3′, turn around: 5′-CTTCCCGTAGAACTTTTTGTTG-3′; GAPDH (Following: 5′-GCACCGTCAAGGCTGAGAAC-3′, switch: 5′-TGGTGAAGACGCCAGTGGA-3′).
Zhang F., Lv M, & He Y. (2024). Identification of a novel disulfideptosis-related gene signature for prognostic implication in lower-grade gliomas. Aging (Albany NY), 16(7), 6054-6067.
+ Open protocol
+ Expand
4

Temporal Expression of Developmental Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols
The WT strain was incubated for 7 days under the optimal regime of 25°C and L:D 12:12 on cellophane-overlaid SDAY (4% glucose, 1% peptone, and 1.5% agar plus 1% yeast extract) plates, which were spread with 100-μl aliquots of a 107-conidia ml−1 suspension for culture initiation. From the end of the 24-h incubation onward, total RNAs were separately extracted daily from three plate cultures using an RNAiso Plus reagent kit (TaKaRa, Dalian, China) and reverse transcribed into cDNAs using a PrimeScript reverse transcription (RT) reagent kit (TaKaRa). Each of the cDNA samples (standardized by dilution) was used as a template to assess transcript levels of brlA and abaA via real-time quantitative PCR (qPCR) with paired primers (see Table S1 in the supplemental material) under the action of SYBR Premix Ex Taq (TaKaRa). The fungal 18S rRNA was used as an internal standard. The threshold cycle (2−ΔΔCT) method (45 (link)) was used to compute the relative transcript level of brlA or abaA in the WT strain on a given day with respect to the standard level at the end of the 24-h incubation.
Zhang A.X., Mouhoumed A.Z., Tong S.M., Ying S.H, & Feng M.G. (2019). BrlA and AbaA Govern Virulence-Required Dimorphic Switch, Conidiation, and Pathogenicity in a Fungal Insect Pathogen. mSystems, 4(4), e00140-19.
+ Open protocol
+ Expand
5

Total RNA Isolation and Purification

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was prepared according to the methods of Ye et al.26 (link),27 . Total RNA was isolated from different tissues using a RNAiso Plus Reagent Kit (Takara Biotechnology, Dalian, China) following the manufacturer’s instructions. The purified RNA was dissolved in RNase-free water, with genomic DNA contamination removed using TURBO DNase I (Promega, Beijing, China). The integrity and purity of the total RNA were checked with a Nanodrop 2000C spectrophotometer (Thermo Scientific, Waltham, Massachusetts) and Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, California). Only the total RNA samples with RIN value ≥ 8 were used for constructing the cDNA library in PacBio or HiSeq sequencing28 (link).
Luo H., Liu H., Zhang J., Hu B., Zhou C., Xiang M., Yang Y., Zhou M., Jing T., Li Z., Zhou X., Lv G., He W., Zeng B., Xiao S., Li Q, & Ye H. (2020). Full-length transcript sequencing accelerates the transcriptome research of Gymnocypris namensis, an iconic fish of the Tibetan Plateau. Scientific Reports, 10, 9668.
+ Open protocol
+ Expand
6

Quantitative Analysis of SP1 Expression in SW480 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was isolated from SW480 cells using the RNAiso Plus reagent kit (Takara). Total RNA (500 ng) was used for reverse transcription using a PrimeScript RT reagent kit (Takara). The resulting complementary DNA was analyzed by qPCR performed with SYBR reagent using the LightCycler 480 PCR system (Roche, Rotkreuz, Switzerland). GAPDH expression was used for normalization. The sequences of used primers were as follows: sp1 (forward: 5′-tggcagcagtaccaatggc-3′, reverse: 5′-ccaggtagtcctgtcagaactt-3′), GAPDH (forward: 5′-ctgggctacactgagcacc-3′, reverse: 5′-aagtggtcgttgagggcaatg-3′).
Ma W., Liu X, & Du W. (2019). Baicalin induces apoptosis in SW480 cells through downregulation of the SP1 transcription factor. Anti-Cancer Drugs, 30(2), 153-158.
+ Open protocol
+ Expand
7

Quantitative Real-Time PCR Protocol for Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted from renal and NRK-52E cells using RNAiso Plus® Reagent Kit according to manufacturer's protocol (Takara Biotechnology Co., Ltd.) and then reverse-transcribed into cDNA using PrimeScript™ RT Reagent kit with DNA Eraser (Takara Biotechnology Co., Ltd.). The cDNA was amplified using SYBR® Premix Ex Taq™ kit (Takara Biotechnology Co., Ltd.). Primer sequences are shown in Table I. The thermocycling conditions of RT PCR were as follows: Initial denaturation at 95˚C for 30 sec, followed by 40 cycles of 95˚C for 5 sec and 60˚C for 30 sec, dissociation at 95˚C for 15 sec and 60˚C for 60 sec and 95˚C for 15 sec. qPCR was subsequently performed using SYBR-Green PCR Master Mix in an ABI prism 7500 Sequence Detection System (Applied Biosystems; Thermo Fisher Scientific, Inc.). The thermocycling conditions of qPCR were as follows: Initial denaturation at 95˚C for 5 min, followed by 40 cycles of 95˚C for 5 sec and 60˚C for 30 sec, dissociation at 95˚C for 15 sec and 60˚C for 60 sec and 95˚C for 15 sec. The 2-ΔΔCq method was used to calculate the fold change for each gene relative to that of GAPDH (35 (link)).
Wang R., Wu G., Dai T., Lang Y., Chi Z., Yang S, & Dong D. (2020). Naringin attenuates renal interstitial fibrosis by regulating the TGF-β/Smad signaling pathway and inflammation. Experimental and Therapeutic Medicine, 21(1), 66.
+ Open protocol
+ Expand
8

Quantifying Gene Expression via RT-qPCR

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNAiso Plus reagent kit (Takara, Kyoto, Japan) was used to extract total RNA according to the manufacture’s requirements. Total RNA was reverse transcribed to cDNA using a Takara Prime Script RT reagent kit (RR047A, Takara, Kyoto, Japan). RT-qPCR was performed using a TB Green PCR kit (RR420A, Takara) with a QuantStudio 6 Flex Real-Time PCR System. The GAPDH gene was used as an endogenous control. The relative gene expression levels were calculated using the 2−ΔΔCT method. The primer sequences used in this study were described in the Supplementary Table S2.
Luo Y.F., Ye X.X., Fang Y.Z., Li M.D., Xia Z.X., Liu J.M., Lin X.S., Huang Z., Zhu X.Q., Huang J.J., Tan D.L., Zhang Y.F., Liu H.P., Zhou J, & Shen Z.C. (2021). mTORC1 Signaling Pathway Mediates Chronic Stress-Induced Synapse Loss in the Hippocampus. Frontiers in Pharmacology, 12, 801234.
+ Open protocol
+ Expand
9

Quantitative Real-time PCR Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
The experimental procedure was performed as described previously (Wen et al., 2018 (link)). Total brain RNA and cDNA synthesis were meausured with an RNAiso Plus Reagent Kit and SYBR Premix Ex Taq Kit, respectively (Takara Biotechnology, Dalian, China). Quantitative real-time PCR was performed using SYBR Green PCR Master Mix and an ABI prim 7500 Sequence Detection System (Applied Biosystems, United States). Primers used for RT-qPCR are shown in Supplementary Table S2.
Du J., Chen X., Zhao Y., Zhao T., Wang D., Chen Z., Wang C., Meng Q., Yao J., Sun H., Liu K, & Wu J. (2022). Characterization of three naturally occurring lignans, sesamol, sesamolin, and sesamin, as potent inhibitors of human cytochrome P450 46A1: Implications for treating excitatory neurotoxicity. Frontiers in Pharmacology, 13, 1046814.
+ Open protocol
+ Expand
10

Quantitative Real-Time PCR for IFN-β, GAPDH, and MERS-N

Check if the same lab product or an alternative is used in the 5 most similar protocols
The culture medium was aspirated, and the transfected cells were lysed with RNAiso Plus reagent kit (Takara Bio). Total cellular RNA was extracted, according to the manufacturer’s protocol. First-strand cDNA synthesis was performed by reverse transcription with Transcriptor First Strand cDNA Synthesis Kit (Roche Diagnostics), according to the manufacturer’s procedures. Quantitative PCR was performed using StepOnePlus Real-Time PCR System (Thermo Fisher Scientific). PCR was run for 40 cycles with an annealing temperature of 60°C. Cycle threshold (CT) values were measured, and relative expression was calculated by ΔCT method normalized with GAPDH as the internal housekeeping gene. Fold expression was calculated by the ΔΔCT method normalized with the mock. The primers were 5′-TTG AAT GGG AGG CTT GAA TA-3′ (forward) and 5′-GCC AGG AGG TTC TCA ACA ATA G-3′ (reverse) for human IFN-β as well as 5′-AAC GTG TCA GTG GTG GAC CTG-3′ (forward) and 5′-AGT GGG TGT CGC TGT TGA AGT-3′ (reverse) for human GAPDH and 5′-CAAAACCTTCCCTAAGAAGGAAAAG-3′ (forward) and 5′-GCTCCTTTGGAGGTTCAGACAT-3′ (reverse) for MERS-N. Data are representative of three independent experiments.
Wong L.Y., Ye Z.W., Lui P.Y., Zheng X., Yuan S., Zhu L., Fung S.Y., Yuen K.S., Siu K.L., Yeung M.L., Cai Z., Woo P.C., Yuen K.Y., Chan C.P, & Jin D.Y. (2020). Middle East Respiratory Syndrome Coronavirus ORF8b Accessory Protein Suppresses Type I IFN Expression by Impeding HSP70-Dependent Activation of IRF3 Kinase IKKε. Journal of immunology (Baltimore, Md. : 1950), 205(6), 1564-1579.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!