Rneasy plus mini kit protocol

Manufactured by Qiagen

The RNeasy Plus Mini Kit is a product designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and fluids. The protocol involves cell lysis, genomic DNA removal, and RNA binding, washing, and elution steps to isolate high-quality RNA for downstream applications.

Automatically generated - may contain errors

Lab products found in correlation

Gotaq qpcr kit, by Promega (1 mentions) Iq5 thermocycler, by Bio-Rad (1 mentions) Cd14 microbead, by Miltenyi Biotec (1 mentions) Anti cd45 percp cy5, by BD (1 mentions) Bme rlt plus buffer, by Qiagen (1 mentions) Automacs technology, by Miltenyi Biotec (1 mentions) Cd19 microbeads, by Miltenyi Biotec (1 mentions) Cd4 microbeads, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

RNeasy Mini Kit (41418 citations) RNeasy Plus Mini Kit (7097 citations) RNeasy Plus kit (1019 citations) RNeasy Mini (299 citations) RNeasy Plus Mini (68 citations) RNeasy protocol (42 citations) RNeasy Mini Protocol (26 citations) RNeasy mini kit protocol (22 citations) RNeasy kit protocol (12 citations) RNeasy kit Mini Kit (2 citations)

2 protocols using rneasy plus mini kit protocol

RNA Extraction and qPCR for Cytokine Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA was extracted from proximal colon samples that were flash frozen at the time of necropsy. Equal sized 5-mm-cubed tissue snips were homogenized using micropestles, and RNA was extracted following the RNeasy Plus Mini Kit protocol (QIAGEN). RNA concentrations were measured using the Nanodrop ND-1000 spectrophotometer and standardized to a concentration of 50 ng/μL. cDNA was obtained by PCR with random primers. A master mix was assembled using reagents from Promega GoTaq qPCR kit and added to the samples. This reaction was run using the following thermal cycler conditions: step 1, 5 min 25 °C; step 2, 20 min 42 °C; step 3, 70 °C; and step 4, 4 °C min—Hold. Interleukin 4 (IL-4) and Interferon gamma (IFNγ) cytokine levels were measured using qPCR on an iQ5 thermocycler (Bio-Rad) with standardization. ANOVAs were performed on 2-ΔΔct data to find the linear fold change in gene expression and are presented as mean fold change of three replicates over levels of the housekeeping gene hypoxanthine-guanine phosphoribosyltransferase (HPRT).

Brooks P.T., Brakel K.A., Bell J.A., Bejcek C.E., Gilpin T., Brudvig J.M, & Mansfield L.S. (2017). Transplanted human fecal microbiota enhanced Guillain Barré syndrome autoantibody responses after Campylobacter jejuni infection in C57BL/6 mice. Microbiome, 5, 92.

+ Open protocol

+ Expand

Isolation and Purification of CD4+ Immune Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fresh PB was subjected to Ficoll density gradient centrifugation. Purified CD4⁺CD19⁻ (total CD4⁺ cells, including CD4⁺ T cells and monocytes), CD4⁺CD14⁻ (CD4⁺ T cells) and CD14⁺ cells (monocytes) were isolated from PBMCs based on positive selection methods employing immunomagnetic beads (CD19 Multisort Kit, CD19 MicroBeads, CD4 MicroBeads and CD14 MicroBeads) and autoMACS technology (Miltenyi Biotec, Bergisch Gladbach, Germany). A flow-chart describing all cell isolation procedures for gene expression analyses is shown in Supplementary Figure 2. To assess the purification process efficacy, 100,000 cells of each sample were analyzed by flow cytometry using anti-CD3 (APC), anti-CD4 (PE), anti-CD14 (FITC) and/or anti-CD45 (PerCP-Cy5.5) antibodies (BD Biosciences, San Jose, CA, USA). Purity of ≥90% was achieved in all samples, except for two MBL subjects in which purities of total CD4⁺ cells were 70%. Purified cells were stored at −80ºC in 1% BME RLT-plus buffer (QIAGEN, Venlo, The Netherlands). RNA was finally extracted following the RNeasy Plus Mini Kit protocol (QIAGEN).

Blanco G., Puiggros A., Sherry B., Nonell L., Calvo X., Puigdecanet E., Chiu P.Y., Kieso Y., Ferrer G., Palacios F., Arnal M., Rodríguez-Rivera M., Gimeno E., Abella E., Rai K.R., Abrisqueta P., Bosch F., Calon A., Ferrer A., Chiorazzi N, & Espinet B. (2021). CLL-like monoclonal B cell lymphocytosis displays an increased inflammatory signature that is reduced in early stage chronic lymphocytic leukemia. Experimental hematology, 95, 68-80.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!