RNA was extracted using a GeneAllR RiboEx total RNA extraction kit (GeneAll Biotechnology, Seoul, Republic of Korea), cDNA reverse-transcription was performed using a Power cDNA synthesis kit (iNtRON Biotechnology, Seongnam, Republic of Korea), and Real-Time RT-PCR was performed in a Step One Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). as previously reported [31 (link)]. The primers used in this study are shown in Supplementary Table S2 .
Geneallr riboex total rna extraction kit
Manufactured by GeneAll
Sourced in Netherlands
The GeneAllR RiboEx total RNA extraction kit is a product designed for the extraction of total RNA from various biological samples. It is a tool used in molecular biology and research laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Power cdna synthesis kit, by iNtRON Biotechnology (5 mentions)
Stepone real time pcr system, by Thermo Fisher Scientific (3 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (2 mentions)
Power sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Qiazol lysis reagent, by Qiagen (1 mentions)
Fast sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
Steponeplus qpcr system, by Thermo Fisher Scientific (1 mentions)
Stepone software v2, by Thermo Fisher Scientific (1 mentions)
8 protocols using geneallr riboex total rna extraction kit
1
RNA Extraction and RT-qPCR Analysis
2
Real-time RT-PCR of Srd5ar2 expression
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Real-time RT-PCR analyses were performed described previously [68 (link)]. Briefly, total RNA from RWPE-1 cells were isolated using a QIAzol lysis reagent (QIAGEN sciences Inc., Venlo, Netherlands) and a GeneAllR RiboEx Total RNA extraction kit (GeneAll Biotechnology, Seoul, Korea). Total RNA was used as a template for first strand cDNA synthesis by a Power cDNA synthesis kit (iNtRON Biotechnology, Seoul, Korea). Step One Plus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) was used for PCR analysis. The primers for Srd5ar2 were Forward 5′ TCC CGC TTG GCC TTT TG-3′ and Reverse 5′- GCC GTT ACC CTC CTT GTT TTC-3′.
3
Skin RNA Extraction and RT-PCR Analysis
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RNA extraction of dorsal skin was performed using the GeneAllR RiboEx total RNA extraction kit (GeneAll Biotechnology, Seoul, South Korea), and Real-time RT-PCR was performed with the Power cDNA synthesis kit (iNtRON Biotechnology, Seongnam, Kyunggi, South Korea), SYBR Green Power Master Mix (Applied Biosystems, Foster City, CA, USA) and the Step One Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) according to the manufacturers’ instructions as previously described (Lim et al., 2016 (link)). The primers used in this study are shown in Table 2
4
RNA Extraction and Real-Time PCR Analysis
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Real-time RT-PCR analyses were performed described previously (Jeong et al., 2015 (link)). Briefly, total RNA from tissues and cells were isolated using a QIAzol lysis reagent (QIAGEN sciences Inc., Venlo, Netherlands) and a GeneAllR RiboEx Total RNA extraction kit (GeneAll Biotechnology, Seoul, Korea). Total RNA was used as a template for first strand cDNA synthesis with a Power cDNA synthesis kit (iNtRON Biotechnology, Seoul, Korea), and Step One Plus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) was used for PCR analyses. The primers used in the experiments are shown in Table 1 .
5
RNA Extraction and Real-Time PCR Protocol
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RNA extraction and real-time PCR were performed with the GeneAllR RiboEx Total RNA Extraction Kit (GeneAll Biotechnology, Seoul, South Korea), Power cDNA Synthesis Kit (iNtRON Biotechnology, Seongnam, South Korea), SYBR Green Power Master Mix (Applied Biosystems, Foster City, CA, United States), and the Step One Real-Time PCR System (Applied Biosystems, Foster City, CA, United States) according to the manufacturers’ instructions as previously described (Kim et al., 2015 (link)). The primers used in this study are provided in Table 1 .
6
Quantifying Gene Expression in Liver and Cell Lines
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Total RNA was extracted using a GeneAllR RiboEx total RNA extraction kit (GeneAll Biotechnology, Seoul, Korea). Newly synthesized complementary DNA (cDNA) from liver or HepG2 cells was amplified using specific primers and the Fast SYBR Green PCR master mix (Applied Biosystems, Foster City, CA, USA). mRNA expression was measured with a StepOnePlus qPCR System and StepOne Software v2.1 (Applied Biosystems, Foster City, CA, USA). The primers used in the experiments are shown in Table 1 .
7
Isolation and Analysis of Spleen Cell RNA in EAE
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The spleen cells were isolated from EAE mice treated with GNPs. Large and small (total) RNA was extracted from splenocytes using GeneAll R RiboEx Total RNA extraction kit (GeneAll Biotechnology, Korea) according to the manufacturer's instructions. The quantity and quality of RNA were evaluated by a Nanodrop spectrophotometer (Thermo Fisher Scienti c, USA). The cDNA synthesis was performed from large RNA in the thermal cycle using miScript Reverse Transcription kit (Parstoos, Iran) according to manufacturer's protocols for the treatment effect on mRNA expression of IL-17 and IFN-cytokines were determined by qPCR. HGPRT was considered as a housekeeping gene. The speci c forward and reverse primers for the evaluation of IL-17 and IFN-genes expression were designed by NCBI primer blast (Table 2). The speci c forward and reverse primers of miR-326-3p were purchased from Pars Genome, Tehran, Iran. Then the qPCR was carried out using the miScript SYBR Green PCR Kit (Pars Genome, Iran) and Qiagen Real-Time PCR system (Applied Biosystem). The RNU6 gene was used as an internal control for normalization. The threshold cycles (CT) were normalized and the relative expression levels were calculated using the 2-ΔΔct method.
8
Isolation and Analysis of Spleen Cell RNA in EAE
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The spleen cells were isolated from EAE mice treated with GNPs. Large and small (total) RNA was extracted from splenocytes using GeneAll R RiboEx Total RNA extraction kit (GeneAll Biotechnology, Korea) according to the manufacturer's instructions. The quantity and quality of RNA were evaluated by a Nanodrop spectrophotometer (Thermo Fisher Scienti c, USA). The cDNA synthesis was performed from large RNA in the thermal cycle using miScript Reverse Transcription kit (Parstoos, Iran) according to manufacturer's protocols for the treatment effect on mRNA expression of IL-17 and IFN-cytokines were determined by qPCR. HGPRT was considered as a housekeeping gene. The speci c forward and reverse primers for the evaluation of IL-17 and IFN-genes expression were designed by NCBI primer blast (Table 2). The speci c forward and reverse primers of miR-326-3p were purchased from Pars Genome, Tehran, Iran. Then the qPCR was carried out using the miScript SYBR Green PCR Kit (Pars Genome, Iran) and Qiagen Real-Time PCR system (Applied Biosystem). The RNU6 gene was used as an internal control for normalization. The threshold cycles (CT) were normalized and the relative expression levels were calculated using the 2-ΔΔct method.
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