Total RNA was isolated via an RNAprep Pure Plant Kit (polysaccharide- and polyphenolic-rich) (Tiangen, Beijing, China). The quality and quantity of the RNA were determined via 1.0% agarose gel electrophoresis and a NanoPhotometer P360 (Implen, Munich, Germany). Reverse transcription was performed with a PrimeScript RT Reagent Kit and gDNA Eraser (Perfect Real Time) (Takara Biotechnology, Dalian, China). The primers used for qRT-PCR are listed in Supplementary Table S1 at JXB online. SYBR Premix Ex Taq (Takara Biotechnology) was used to perform quantitative real-time PCR (qRT-PCR). The thermal cycling conditions for qRT-PCR were as follows: predenaturation at 95 °C for 10 min followed by 40 cycles of denaturation at 95 °C for 5 s, annealing at 60 °C for 30 s, and extension. Each sample was replicated three times.
Gdna eraser perfect real time
Manufactured by Takara Bio
Sourced in China, Japan
The GDNA Eraser (Perfect Real Time) is a laboratory equipment designed to remove genomic DNA (gDNA) from RNA samples. It is a tool used in molecular biology applications to ensure the purity of RNA preparations, which is essential for accurate downstream analysis such as real-time PCR.
Automatically generated - may contain errors
Lab products found in correlation
Primescript rt reagent kit, by Takara Bio (33 mentions)
Trizol reagent, by Thermo Fisher Scientific (8 mentions)
Rnaiso plus, by Takara Bio (6 mentions)
Sybr premix ex taq, by Takara Bio (3 mentions)
Sybr premix ex taq tli rnaseh plus, by Takara Bio (3 mentions)
Nanophotometer p360, by Implen (2 mentions)
Rnaprep pure plant kit polysaccharide and polyphenolic rich, by Tiangen Biotech (2 mentions)
Primerscript rt reagent kit, by Takara Bio (2 mentions)
Sybr premix ex taq kit, by Takara Bio (2 mentions)
Sybr green pcr kit, by Takara Bio (2 mentions)
Rneasy micro kit, by Qiagen (2 mentions)
Emeraldamp max pcr master mix, by Takara Bio (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Tb green premix ex taq 2 tli rnaseh plus, by Takara Bio (2 mentions)
Biophotometer, by Eppendorf (1 mentions)
Axyprep multisource total rna miniprep kit, by Corning (1 mentions)
Sqrt pcr rnaiso plus, by Takara Bio (1 mentions)
Smartspec plus spectrophotometer, by Bio-Rad (1 mentions)
Rnaout 2.0 kit, by Tiandz (1 mentions)
Abi 7900ht sequence detection system, by Thermo Fisher Scientific (1 mentions)
46 protocols using gdna eraser perfect real time
1
RNA Extraction and qRT-PCR Analysis
2
Developmental Expression Profiles of LeMet and LeKr‐h1
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Total RNA was extracted from the whole body or different tissues of female adults using AxyPrep Multisource Total RNA Miniprep Kit (Axygen) according to the manufacturer's protocol. To investigate the developmental expression profiles of LeMet and LeKr‐h1, individuals were collected from eggs and nymphs (1st, 2nd, 3rd, and 4th instars), females and males within 1 week after emergence, and adult females on different days after emergence. Tissues of adult females within 1 week after emergence (including head, thorax, midgut, ovary, and fat body) were dissected in phosphate‐buffered saline (PBS). All samples were frozen immediately in liquid nitrogen and stored at −80°C until RNA extraction. The extracted RNA was instantly dissolved in TE buffer and checked for quality, concentration, and purity at an absorbance ratio of optical density (OD) 260/280 (1.8–2.1) on a BioPhotometer (Eppendorf). Each experiment contained 30 individuals and was performed in at least three biological replicates. Finally, first‐strand cDNA was synthesized from 1 µg total RNA using a PrimeScript™ RT reagent kit with gDNA Eraser (Perfect Real Time) (Takara), following the manufacturer's protocol. The synthesized first‐strand cDNA was stored at –20°C until further processing.
3
Gill Tissue RNA Extraction and qRT-PCR
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Total RNA was isolated from gill tissue using sqRT-PCR RNAiso Plus (TaKaRa, Dalian, China). The cDNA was synthesized using the Perfect Real Time version of the PrimerScriptTM RT reagent kit with gDNA Eraser (Perfect Real Time) (TaKaRa,) according to manufacturer’s instructions. Then, sq-RT-PCR were chosen to analyze genes, and performed in a total reaction volume of 25 μl according to the manufacturers’ instructions. The S. paramamosain beta-actin gene and 18S ribosomal RNA gene were selected as the internal control. Primers used in this study are listed in Additional file 1 : Table S1.
4
RNA Extraction and cDNA Synthesis
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Total RNA was isolated from various samples with an RNAout 2.0 kit (Tiandz, Beijing, China) according to the manufacturer’s instructions. To remove trace DNA from samples, total RNA extractions were treated with RNase free DNase I. The RNA integrity was detected using 2% agarose gels. A Bio-RAD smart spec™plus spectrophotometer was used to determine the RNA purity. cDNA was synthesised with a PrimeScript™ RT reagent kit and gDNA Eraser (Perfect Real Time; TaKaRa, Dalian, China).
5
Quantitative Analysis of MLL4 Expression
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RNA was reversely transcribed into cDNA using a PrimeScript RT reagent kit with a gDNA Eraser (Perfect Real Time) (TaKaRa Biotechnology, Dalian, China) according to the manufacturer’s instructions. Reactions of real-time PCR were performed on ABI 7900 HT Sequence Detection System (Applied Biosystems) using TaqMan Gene Expression Master Mix (4369016, Applied Biosystems) according to the manufacturer’s instructions. TaqMan Gene Expression Assay (Hs00207065_m1, Applied Biosystems) was used to detect MLL4 expression level. MLL4 values are shown as relative expression levels to 18s (4319413, Applied Biosystems) using formula 2-ΔCt (MLL4-18s)*105.
6
RNA Extraction and cDNA Synthesis
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The total RNA of persimmon was isolated following the hot borate method (Wan and Wilkins, 1994 (link)). The total RNA of the Arabidopsis and tomato were extracted using a TransZol Up Plus RNA Kit (Transgen Biotech, Beijing, China). The first-strand cDNA was synthesized from 1 μg of RNA using the PrimeScriptTM RT Reagent Kit with the gDNA Eraser (Perfect Real Time; TaKaRa, Dalian, China) following the manufacturer’s protocol. The synthesized cDNA was diluted 10-fold for persimmon and tomato samples or 20-fold for Arabidopsis leaves for the following qPCR analysis.
7
Quantitative RT-PCR Analysis of CsBPC Expression
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Total RNA was extracted using an RNA prep pure Plant Kit (TANGEN) and first-strand cDNA was synthesized using a PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TaKaRa) according to the manufacturer’s instructions. Quantitative RT-PCR was performed following the instructions of the SYBR® Premix Ex Taq™ Kit (TaKaRa) on a Mx3000P real-time PCR instrument (Agilent). The experiments were performed with three biological replicates, each with three plants. Relative gene expression was calculated using the 2−△△Ct method [51 (link)]. The CsBPC specific primers used for expression pattern analysis in different tissues and under abiotic stress and hormone treatments are shown in Additional file 2. S1 .
8
Quantifying HOXA11-AS in OSCC
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Total cellular RNA was isolated from primary OSCC tissues using TRIzol reagent (Invitrogen) and reverse-transcribed using the Prime Script RT reagent kit together with gDNA Eraser (Perfect Real Time; Takara, Kyoto, Japan) in accordance with the manufacturer’s instructions. HOXA11-AS expression was analyzed using qRT-PCR, with reactions performed in triplicate using a SYBR Green PCR kit (Takara). Glyceraldehyde3-phosphate dehydrogenase (GAPDH) mRNA was used as the internal control. The primer sets are listed in Table 1 .
9
RNA Isolation and RT-PCR Analysis
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Total RNAs were isolated using an RNeasy Micro Kit (QIAGEN, Tokyo, Japan). cDNAs were synthesized using PrimeScript® RT reagent Kit with gDNA Eraser Perfect Real Time (TaKaRa BIO, Otsu, Japan). RT-PCR was performed using EmeraldAmp® MAXPCR Master Mix (TaKaRa). Primer sequences are listed in the Supplemental Table 1 .
10
Quantifying OSCC RNA Expression
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Total cellular RNA was isolated from primary OSCC tissues using TRIzol reagent (Invitrogen) and reverse-transcribed using the Prime Script RT reagent kit together with gDNA Eraser (Perfect Real Time; Takara, Kyoto, Japan) in accordance with the manufacturer’s instructions. RNA expression was analyzed using qRT-PCR, with reactions performed in triplicate using a SYBR Green PCR kit (Takara). Glyceraldehyde3-phosphate dehydrogenase (GAPDH) mRNA was used as the internal control. The primer sets were listed in Table 1 .
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