BM MSCs or MPCs were sorted, and RNA was extracted using RNeasy MicroKit (Qiagen) according to the manufacturer's recommendations. cDNA was prepared using SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Takara Bio). The cDNA quality was examined on the Agilent TapeStation system using a High Sensitivity D5000 ScreenTape (Agilent). One ng cDNA was used for library preparation using Nextera XT DNA Library Preparation Kit (FC-131-1024 & FC-131-1096, Illumina). The yield and quality of the amplified libraries were analyzed using Qubit (Thermo Fisher Scientific) and the Agilent TapeStation system. The indexed cDNA libraries were normalized, combined, and sequenced on the Illumina Nextseq 550 or 2000 for a 75-cycle v2 or 130-cycle P2 sequencing run generating 75 bp single-end or pair-end reads. See supplemental Data for data analysis.
Agilent tapestation system
Manufactured by Agilent Technologies
Sourced in United States, United Kingdom
The Agilent TapeStation system is a versatile and automated platform for the analysis of nucleic acid samples. It provides rapid and reliable sample quality assessment and quantification for a wide range of applications, including next-generation sequencing, PCR, and genomic research.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (7 mentions)
Qubit 2.0 fluorometer, by Thermo Fisher Scientific (5 mentions)
Turbo dna free kit, by Thermo Fisher Scientific (4 mentions)
Quantus fluorometer, by Promega (3 mentions)
Direct zol rna miniprep kit, by Zymo Research (3 mentions)
Turbo dnase, by Thermo Fisher Scientific (2 mentions)
Onetouch 2 instrument, by Thermo Fisher Scientific (2 mentions)
Ion proton instrument, by Thermo Fisher Scientific (2 mentions)
Rneasy mini kit, by Qiagen (2 mentions)
Hiseq 4000, by Illumina (2 mentions)
Nanodrop spectrophotometer, by Agilent Technologies (2 mentions)
Nextseq 500, by Illumina (2 mentions)
Epitect pcr control dna set, by Qiagen (2 mentions)
Hotstartaq dna polymerase, by Promega (2 mentions)
Epitect bisulfite kit, by Qiagen (2 mentions)
Dntps, by Promega (2 mentions)
Novaseq 6000, by Illumina (2 mentions)
Hiseq 2500 sequencer, by Illumina (2 mentions)
Nebnext rna first and second strand synthesis module, by New England Biolabs (2 mentions)
Rnaprotect reagent, by Qiagen (2 mentions)
33 protocols using agilent tapestation system
1
Transcriptome Profiling of BM-Derived Cells
2
RNA Extraction from S. lugdunensis Mutants
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RNAs were extracted from cells of S. lugdunensis N920143 and ΔagrA mutant cultured in BHI harvested in the late-exponential phase (OD600 of 1) as previously described (16 (link)). Following incubation of the cell pellets for 12 h at −80°C, RNAs were extracted with the Direct-zol RNA miniprep kit (Zymo Research, Irvine, CA, USA) and residual DNA was removed using Turbo DNase (ThermoFisher) according to the manufacturer’s instructions. RNAs integrities and quantifications were measured by the Agilent TapeStation system (Agilent, Santa Clara, CA, USA). Four independent samples for each strain were used.
3
Ion Exome Sequencing Protocol
4
Ion Ampliseq-based Gene Panel Sequencing
5
Extraction and Sequencing of Circulating RNA
6
Single-cell RNA-seq of Chicken Pituitary Cells
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Pituitary cell dispersion was carried out as described in our previous studies (Meng et al., 2014 (link)). Briefly, anterior pituitaries were separated from adult male and female chickens (1-year-old). Then, six male/female pituitaries were pooled and washed in 1 × Hank’s balanced salt solution (HBSS) thrice. These pituitaries were treated by 0.25% trypsin solution for 30 min at 37°C. Dissociated cells were filtered through a 40-μm cell strainer and subject to centrifugation at 500 g for 3 min. Then the cell pellet was re-suspended in a solution containing 0.1% bovine serum albumin (BSA) in 1 × PBS solution (calcium- and magnesium-free). Cell viability and numbers were estimated by trypan blue staining. Then, pituitary cells were loaded onto the Chromium system (10x Genomics 3′ GEX V2) to produce cDNA libraries according to the Chromium Single Cell 3′ Reagents Kits V2 User guide (10x Genomics Inc., Pleasanton, CA, United States). The cDNA libraries were quantified by Agilent Tapestation System (Agilent Technologies, Santa Clara, CA, United States) and sequenced by BGI500 (BGI, Shenzhen, China).
7
RNA Extraction from Spinal Cord Tissue
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Methods were described in our previous study.18 (link) Briefly, 14 days after the model establishment, both model and sham groups of rats were anesthetized deeply using sodium pentobarbital at a dosage of 40 mg/kg (i.p.). The animals were then perfused with 0.9% saline with 4 °C. Then the ipsilateral SCDH tissues were harvested and immediately preserved in RNAlater solution (#AM7020, Invitrogen, USA). Trizol reagent (#15596018, Thermo Fisher, USA) was used for total RNA extraction. The concentrations and the purities of tissue samples was examined using Nanodrop Spectrophotometer (NanoDrop One, Thermo Fisher, USA). RNA integrity number (RIN) was then determined by Agilent TapeStation System (Agilent Technologies, USA).
8
Isolating RNA from Rat Skin Samples
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CPIP or control group rats were anesthetized deeply using sodium pentobarbital at dosage of 40 mg/kg. The animals were then perfused with 0.9% saline with 4°C. Then, the ipsilateral hindpaw skin was harvested and immediately preserved in RNAlater solution (Thermo Fisher, USA). TRIzol reagent (Thermo Fisher, USA) with DNase I was used for total RNA extraction. The concentrations and the purities of skin samples were evaluated using the NanoDrop Spectrophotometer (NanoDrop Products, USA). RNA integrity number (RIN) was determined by the Agilent TapeStation System (Agilent Technologies, USA).
9
Total RNA Extraction from WJ-MSCs
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Total RNA was extracted from WJ-MSCs samples using QIAGEN RNeasy Mini-Kit (QIAGEN, Hilden, Germany). RNA from all samples was run on an Agilent TapeStation System (Agilent Technologies, Santa Clara, CA, USA) to assess quality, and to estimate RNA concentrations, RNA was quantitated by Promega QuantiFluor Dye System on Quantus Fluorometer (Promega).
10
Total RNA Extraction and Quantification
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Total RNA was extracted from Cardio-0007 and Cardio-0007 Luc+ samples using the QIAGEN RNeasy Mini-Kit (QIAGEN, Hilden, Germany). RNA from all samples was run on an Agilent TapeStation System (Agilent Technologies, USA) to assess quality, and to estimate RNA concentrations, RNA was quantitated by Promega QuantiFluor Dye System on Quantus Fluorometer (Promega, USA).
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