The total RNA of cADMSCs was extracted by using Trizol reagent (Takara, Japan) according to the manufacturer’s instructions. Reverse Transcriptase Reagent kit (Thermo Scientific) was used to reverse transcript RNA into cDNA according to the manufacturer’s instructions. QRT-PCR was performed in the CFX96 Real-Time PCR system, and the QRT-PCR procedures were described as follows: pre-denaturation at 94 °C for 5 min, following 39 cycles for 30 s at 94 °C, annealing for 30 s at 58 °C and 30 s at 70 °C for extending. Gapdh was used as the loading control. Comparative CT values from QRT-PCR were used to measure relative gene expression. Primers are listed in Table S1 .
Reverse transcriptase reagent kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Reverse Transcriptase Reagent kit is a laboratory tool used for the synthesis of complementary DNA (cDNA) from RNA templates. It contains the necessary components, including reverse transcriptase enzyme, required for the reverse transcription process.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Takara Bio (3 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Cfx96 qrt pcr detection system, by Bio-Rad (1 mentions)
Rnaiso, by Takara Bio (1 mentions)
Cell culture flask, by SPL Life Sciences (1 mentions)
Trypan blue solution 0.4, by Merck Group (1 mentions)
3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions)
Well plates, by SPL Life Sciences (1 mentions)
5 protocols using reverse transcriptase reagent kit
1
Quantifying Gene Expression in cADMSCs
2
Goat Testis RNA Extraction and qRT-PCR
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Total RNA was extracted from goat testisand cultured SSCs using RNAiso (TaKaRa, Biotech. Co. Ltd., Dalian, China). Total RNA was reverse transcribed into cDNAs using the Reverse Transcriptase reagent kit according to the manufacturer's instructions (Thermo Scientific, FL33407, USA). QRT-PCR was performed on a CFX96 QRT-PCR detection system (Bio-Rad, CA 94547, USA) according to the instructions for the BioEasy SYBR Green I RT-qPCR kit (Bioer Co. Ltd., Hangzhou, China). The QRT-PCR was performed as described previously [26 (link)]. The relative expression levels of the target genes were normalized to Gapdh expression for each sample. The relative expression levels were calculated using 2−ΔΔCt. The primers for the validated mRNAs are listed in S2 Text.
3
Quantitative Real-Time PCR Analysis of cADMSCs
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The total RNA of cADMSCs was extracted using Trizol reagent (Takara, Japan) according to the manufacturer’s instructions. Reverse Transcriptase Reagent kit (Thermo Scientific) was used according to the manufacturer’s instructions. Quantitative real-time PCR (QPCR) was carried out with the CFX96 Real-Time PCR system as follows: pre-denaturation at 94 °C for 5 min, following 39 cycles for 30 s at 94 °C, annealing for 30 s at 58 °C and 30 s at 70 °C for extending. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control. Comparative CT-values from QPCR were used to measure relative gene expression. Primers are listed in Supplementary Table 2 .
4
Pistachio Pollens and Stem Cell Interactions
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The mature PGs of Pistachio (Pistacia vera L.) were collected from male flowers in early spring in Qazi Jahan Pistachio gardens, Iran. The hAD-MSCs were obtained from the Iranian Biological Resource Center (Tehran, Iran). Trypan blue solution (0.4%) and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Poole, UK). TRIzol reagent and reverse transcriptase reagent kit were bought from Thermo Fisher Scientific (Waltham, MA, USA). Maxima SYBER Green qPCR master mix was obtained from BioFACT (Daejeon, Korea). Cell culture flasks, well plates, and pipettes were purchased from SPL Life Sciences (Pocheon, South Korea). All cell culture media and components were provided from Gibco, Life Technologies (Paisley, UK). The oligonucleotide primer sequences were received from Takapo Zist Company (Tehran, Iran). Other chemicals were supplied from Merck (Kenilworth, NJ, USA).
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The mature PGs of Pistachio (Pistacia vera L.) were collected from male flowers in early spring in Qazi Jahan Pistachio gardens, Iran. The hAD-MSCs were obtained from the Iranian Biological Resource Center (Tehran, Iran). Trypan blue solution (0.4%) and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Poole, UK). TRIzol reagent and reverse transcriptase reagent kit were bought from Thermo Fisher Scientific (Waltham, MA, USA). Maxima SYBER Green qPCR master mix was obtained from BioFACT (Daejeon, Korea). Cell culture flasks, well plates, and pipettes were purchased from SPL Life Sciences (Pocheon, South Korea). All cell culture media and components were provided from Gibco, Life Technologies (Paisley, UK). The oligonucleotide primer sequences were received from Takapo Zist Company (Tehran, Iran). Other chemicals were supplied from Merck (Kenilworth, NJ, USA).
5
Quantitative Real-Time PCR for Gene Expression
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According to the manufacturer’s instructions, the total RNA was extracted from the ADMSCs by TRIzol reagent (Takara, Japan), and a reverse transcriptase reagent kit (Thermo Fisher Scientific) was used. Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out using a CFX96 Real-Time polymerase chain reaction (PCR) system as follows: predenaturation at 94°C for 5 min, followed by 39 cycles for 30 s at 94°C, annealing for 30 s at 58°C and 30 s at 70°C for extension. β-Actin was used as an internal control. The comparative CT values from the qRT-PCR were used to measure the relative gene expression (Wei et al., 2016 (link); Zhu et al., 2021 ). The primers are listed in Supplementary Table 1 .
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