RNeasy 96 BioRobot 8000 Kit (12)

Protocol tips

Upstream tips	Protocol tips	Downstream tips

Publication protocol

Preparation of the prey mRNA library: An RNA library for mRNA display was created from the poly(A)+ mRNAs extracted from the spleens of 6- to 8-week-old C57BL/6 mice, according to a previously described method7,13. First, randomly primed reverse transcription (RT) of poly(A)+ mRNAs were subjected to ligation-mediated amplification14 and multi-step PCRs to create cDNA constructs for in vitro expression. The resulting PCR products (SP6-Ω-T7-Flagment-Kpn1-FLAG-A{8}) were purified with a QIAquick PCR Purification Kit (Qiagen, Germany) and transcribed into mRNA with a RiboMAX Large Scale RNA Production System-SP6 (Promega, WI, USA) and an m7G(5′)ppp(5′)G RNA Cap Structure Analog (Ambion, Life Technologies, CA, USA). After purification of the transcribed mRNAs using an RNeasy 96 BioRobot 8000 Kit (Qiagen), PEG Puro spacer was ligated to the 3′ends of mRNAs using T4 RNA ligase (Promega) and the RNA was purified again.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing mRNA / Ribonucleoprotein isolation / purification mRNA using RNeasy 96 BioRobot 8000 Kit (12) from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for RNeasy 96 BioRobot 8000 Kit (12) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing mRNA / Ribonucleoprotein isolation / purification mRNA using RNeasy 96 BioRobot 8000 Kit (12) from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.