Total RNA was isolated from mouse brains, using TRIzol (GRISP, Ref. No GB23.0100; Porto, Portugal) and the RNeasy Mini Kit (Machery-Nagel; Dueren, Germany) from blood using the NucleoSpin® RNA Blood (Machery-Nagel; Dueren, Germany) and from cell suspension using the NucleoSpin® RNA XS (Machery-Nagel, Ref. No 12733391; Dueren, Germany). Total RNA was retrotranscribed to cDNA (Transcriptor First Strand cDNA Synthesis Kit, ThermoFisher LTI, Ref. No 18080-051; Bleiswijk, The Netherlands) for PCR with Power SYBR Green PCR master mix (BioRad, Ref. No 1725124; Madrid, Spain). Transcript number was calculated from the threshold cycle (Ct) of each gene with a 2−ΔΔCT method (relative number), normalized to ArbP0 or GADPH, and expressed as fold induction of animals used as controls. The primers used in this study are listed below in Table 1 .
Nucleospin rna blood
Manufactured by Macherey-Nagel
Sourced in Germany
NucleoSpin RNA Blood is a silica-membrane-based RNA extraction kit designed for the efficient isolation of high-quality total RNA from whole blood samples. The kit utilizes a simple and fast protocol to extract RNA, which can be used for downstream applications such as qRT-PCR, RT-PCR, and Northern blotting.
Automatically generated - may contain errors
Lab products found in correlation
Nucleospin rna xs, by Macherey-Nagel (1 mentions)
Rneasy mini kit, by Macherey-Nagel (1 mentions)
Power sybr green pcr master mix, by Bio-Rad (1 mentions)
Rneasy minelute cleanup kit, by Qiagen (1 mentions)
Rnaiso plus, by Takara Bio (1 mentions)
Dnase, by Qiagen (1 mentions)
Quantitect reverse transcription kit, by Qiagen (1 mentions)
3 protocols using nucleospin rna blood
1
RNA Extraction and qRT-PCR Analysis
2
Total RNA Extraction from Leukocytes and Spleen
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Total RNA was extracted from peripheral leukocytes, splenocytes, and spleen tissues as described previously using the acid guanidine thiocyanate-phenolchloroform method and silica membrane column-based purification [12] (link). Briefly, cells or tissues were homogenized in RNAiso Plus (Takara Bio Inc.), and the total RNA was isolated. The RNA was treated with DNase (Qiagen, Valencia, CA, USA) in the aqueous phase and further purified using the RNeasy MinElute Cleanup Kit (Qiagen) according to the manufacturer's instructions.
Total RNA (0.2 -0.4 mL) of mouse whole blood was isolated using NucleoSpin RNA Blood (Macherey-Nagel GmbH & Co.) according to the manufacturer's instructions with on-column DNA digestion.
The quantity and purity of RNA were evaluated at 230, 260, 280, and 320 nm using an Ultrospec 2000 spectrometer (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).
Total RNA (0.2 -0.4 mL) of mouse whole blood was isolated using NucleoSpin RNA Blood (Macherey-Nagel GmbH & Co.) according to the manufacturer's instructions with on-column DNA digestion.
The quantity and purity of RNA were evaluated at 230, 260, 280, and 320 nm using an Ultrospec 2000 spectrometer (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).
3
Endometrial RNA Extraction and cDNA Synthesis
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Total RNA was extracted from endometrial cells and used for cDNA synthesis using established protocols in our laboratory [26 (link)]. Briefly, RNA was isolated using a commercial kit (NucleoSpin RNA Blood, Cat No. 40200, Macherey-Nagel GmBH&Co. KG, Büren, Germany).
All RNA samples were quantified by spectrophotometry (#ND-1000, Nanodrop Technology Inc.; Wilmington, DE, USA), and the purification of RNA with A260/A280 ratio was between 1.8 and 2.0. Complementary DNA (cDNA) was synthesized from 150 ng RNA using a QuantiTect Reverse Transcription kit (Qiagen, Hilden, Germany, Cat No. 205314), as described by Dirandeh et al. (2021) [3 (link)].
All RNA samples were quantified by spectrophotometry (#ND-1000, Nanodrop Technology Inc.; Wilmington, DE, USA), and the purification of RNA with A260/A280 ratio was between 1.8 and 2.0. Complementary DNA (cDNA) was synthesized from 150 ng RNA using a QuantiTect Reverse Transcription kit (Qiagen, Hilden, Germany, Cat No. 205314), as described by Dirandeh et al. (2021) [3 (link)].
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