Total RNA was isolated from cells or thymus tissues using RNeasy Plus Spin Column Kits or RNeasy Plus Universal Mini Kits (Qiagen, Valencia, CA, USA) according to the manufacturer’s instructions. Poly A+ RNA or total RNA from normal human tissues was purchased from Clontech (Mountain View, CA, USA). Complementary DNAs were synthesized using the SuperScript Preamplification System (Invitrogen, Carlsbad, CA, USA). qPCR was conducted with SYBR Green Master Mix on a LightCycler 480 (Roche, Basel, Switzerland). Primer sequences are shown in Supplementary Table S1 .
Rneasy plus spin column kits
Manufactured by Qiagen
Sourced in United States
The RNeasy Plus Spin Column Kits are designed for the purification of total RNA from various sample types. The kits utilize spin columns to efficiently capture and elute RNA, providing a simple and effective method for RNA extraction.
Automatically generated - may contain errors
2 protocols using rneasy plus spin column kits
1
RNA Isolation and qPCR Analysis
2
RNA Extraction and qRT-PCR Analysis of DHT-Treated LNCaP Cells
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Extraction of total RNA from DHT-treated and untreated LNCaP cells was performed using the RNeasy plus spin column kits (Qiagen, Valencia, CA, USA) according to the manufacturer protocol. Complementary DNAs (cDNAs) were prepared with the iScript cDNA Synthesis Kit (Bio-Rad Laboratories, CA, USA). qRT-PCR analysis was performed using the SYBR Green Master CFX Connect Real-Time PCR System (Bio-Rad Laboratories, CA, USA).28 (link),29 (link) Preparation of complementary DNAs (cDNAs) and qRT-PCR was conducted by the iScript cDNA Synthesis Kit (Bio-Rad Laboratories, CA, USA) and the SYBR Green Master on a CFX Connect Real-Time PCR System (Bio-Rad Laboratories, CA, USA), respectively.29 (link),30 (link) Expression of eRNA transcripts was calculated by the delta–delta Ct method with β-actin expression used as a normalizing internal control. The PCR primer sets used in the study are listed in Supplementary Table 1
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