Total RNA from transplanted tumor tissues was isolated with Multisource Total RNA Miniprep Kit (Axygen), and 1 μg was reverse-transcribed with random hexamer primer using the First-Strand cDNA Synthesis SuperMix (TransGen). According to the manufacturer's instructions, 5 ng cDNA was amplified in triplicate in a reaction volume of 10 μl with the StepOne Plus Real-time PCR System (Applied Biosystems) and StepOne Software v2.2.2 (Applied Biosystems). The expression level was normalized against the geometric mean of β-actin. Relative mRNA expression was calculated using the 2−ΔΔCT method.
Multisource total rna miniprep kit
Manufactured by Corning
Sourced in United States, China
The Multisource Total RNA Miniprep Kit is a laboratory equipment product designed for the isolation and purification of total RNA from a variety of sample types. The kit utilizes a silica-based membrane technology to efficiently capture and purify RNA, providing a reliable and consistent method for RNA extraction.
Automatically generated - may contain errors
Lab products found in correlation
Sybr premix ex taq 2, by Takara Bio (4 mentions)
Primescript rt master mix, by Takara Bio (3 mentions)
Abi prism 7000, by Thermo Fisher Scientific (3 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (2 mentions)
First strand cdna synthesis supermix, by Transgene (1 mentions)
Stepone software v2, by Thermo Fisher Scientific (1 mentions)
Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
A reverse transcription kit, by Promega (1 mentions)
Real time pcr system, by Bio-Rad (1 mentions)
Oligo dt beads, by Illumina (1 mentions)
Sequencing adapter, by Illumina (1 mentions)
Qiaquick pcr extraction kit, by Qiagen (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
Sybr green pcr master mix, by Takara Bio (1 mentions)
Transscript fly first strand cdna synthesis supermix, by Transgene (1 mentions)
Rtaq dna polymerase, by Takara Bio (1 mentions)
Dnase 1, by Thermo Fisher Scientific (1 mentions)
M mlv reverse transcriptase, by Promega (1 mentions)
Cdna reverse transcription kit, by Takara Bio (1 mentions)
Primescript rt pcr kit, by Takara Bio (1 mentions)
17 protocols using multisource total rna miniprep kit
1
Quantitative Real-Time PCR Analysis of Tumor RNA
2
Quantifying Aass Gene Expression
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RNA was isolated from different tissues of WT (C57/B6) mice using a Multisource Total RNA Miniprep Kit (Axygen). A reverse-transcription kit (Promega) was used to reverse-transcribe RNA in a 20-µl reaction mixture. Quantification of Aass gene expression was performed using a real-time PCR system (Bio-Rad) in quadruplicate. Gapdh was amplified as the internal control.
3
Quantitative Real-Time PCR for Gene Expression
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Total RNA was isolated from tissue or cultured cells using Multisource Total RNA Miniprep Kit (Axygen, 365), and then used for cDNA synthesis (PrimeScript™RT Master Mix, Takara, RR036Q) and amplification by real-time PCR according to the manufacturer’s instructions (SYBR Premix Ex Taq™ II, Takara, RR820A). Relative gene expression quantification was based on the comparative threshold cycle method (2−ΔΔCt) using β-actin as endogenous control gene. The primer sequences were provided in Additional file 1 : Table S2.
4
RNA Extraction and qRT-PCR Analysis
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RNA was extracted using a Multisource Total RNA Miniprep Kit (AXYGEN, CA, USA) and qRT-PCR was performed using commercially available primers and SYBR Premix Ex Taq II (DRR081; Takara Biotechnology Co. Ltd., Dalian, China). Fluorescence for each cycle was quantitatively analyzed using the ABI Prism 7000 sequence detection system (Life Technologies). The results were reported as relative expression, normalized to β-actin housekeeping gene as an endogenous control and expressed in arbitrary units. The sequence of the primers used is listed in Table S3 .
5
RNA Extraction and qRT-PCR Analysis
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RNA was extracted using a multisource total RNA miniprep kit (AXYGEN Biosciences, Hangzhou, China) according to manufacturer's instructions. qRT-PCR was performed as described previously. Primers are listed in Supplementary Table 2 .
6
Time-course RNA-seq of Infected Neurons
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Total RNA from primary cultured neurons after 1,3,5,7,10 days of infection was extracted using Multisource Total RNA Miniprep Kit (Axygen, 365). RNA quality was performed using Agilent 2100 Bioanalyzer (Agilent Technologies, USA) and checked using RNase free agarose gel electrophoresis. After qualification, total mRNA was enriched by Oligo (dT) beads (Epicentre, USA). RNA was fragmented into short fragments and reverse transcript into cDNA with random primers. Second-strand cDNA were synthesized and the cDNA fragments were purified with QiaQuick PCR extraction kit (Qiagen, Netherlands), end repaired, poly A added, and ligated to Illumina sequencing adapters. Selecting ligation products with proper size by PCR and agarose gel electrophoresis, and then sequencing was performed by Gene Denovo Biotechnology Co. (Guangzhou, China) on Illumina HiSeq2500.
7
Quantification of Penile Gene Expression
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Total RNA was obtained from the corpus cavernosum using a Multisource Total RNA Miniprep Kit (AXYGEN, Union City, CA, USA) according to the manufacturer’s instructions. Reverse transcription and real-time PCR were conducted using PrimeScript RT Master Mix and SYBR Green PCR Master Mix (TaKaRa, Dalian, Liaoning, China), as described in our previous studies [23 (link)]. The mRNA expression levels of the examined genes relative to β-actin were calculated using the 2-ΔCt method. The primer sequences used for hKLK1, rat tissue KLK1 (rKLK1), eNOS, nNOS, COX-2, PTGIS and β-actin were shown in Table 1 .
8
Hyphopodium Differentiation Gene Expression
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Expression levels of genes involved in the hyphopodium differentiation were measured after the test strains were grown on MM plates covered with cellophane membrane for 2 days to induce hyphopodium development. Total RNA was extracted from the mycelium using the Multisource Total RNA Miniprep Kit (Axygen). RT‐qPCR was performed under the following conditions: an initial 95 °C denaturation step for 10 min followed by 40 cycles of 95 °C for 15 s and 60 °C for 1 min. The transcription levels of the target genes relative to the constitutively expressed elongation factor 1‐α of Verticillium dahliae were quantified using the 2−∆∆Ct method from three independent experiments (Livak & Schmittgen, 2001 (link)). Unpaired Student's t tests were performed to determine statistical differences. Primers used for expression profiling are listed in Table S1 .
9
Total RNA Isolation and cDNA Synthesis
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Total RNA was isolated using a Multisource Total RNA Miniprep kit (Axygen Scientific, CA, USA) according to the manufacturer’s instructions. First-strand cDNA synthesis was conducted with 5 μg of total RNA by using a TransScript Fly First-strand cDNA Synthesis SuperMix (Transgen Biotech, Shanghai, China).
10
RNA Extraction and cDNA Synthesis
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Total cellular RNA was extracted using a Multisource Total RNA Miniprep Kit (Axygen, Tewksbury, MA, USA). RNA was treated with DNase I (Invitrogen) and reverse-transcribed using random primers (hexadeoxynucleotides) (Promega, Madison, WI, USA) and Moloney Murine Leukemia Virus (MMLV) Reverse Transcriptase (Promega). PCR was performed using rTaq DNA polymerase (TaKaRa, Tokyo, Japan). The primer sequences are listed in the Table S1 .
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