Therneasy kit

Manufactured by Qiagen

Sourced in United States, Germany

The RNeasy kit is a laboratory equipment product designed for the purification and isolation of total RNA from a variety of sample types. It utilizes a silica-membrane-based technology to efficiently capture and purify RNA molecules.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

TheRNeasy Mini Kit (11 citations) TheRNeasy plus micro kit (2 citations)

6 protocols using therneasy kit

qPCR Analysis of Gene Expression

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

Total mRNA from cells and tissue specimens was isolated using the
RNeasy kit (Qiagen). mRNA was reverse transcribed to cDNA using the high
capacity cDNA reverse transcription kit (Thermo Fisher). qPCR was performed
using the StepOne Real-Time PCR system (Applied Biosystems) and qPCR master
mix (Kapa Biosystems) with standard cycling parameters. TaqMan qPCR primer
sets for human MPC1, MPC2, and
KLK3 (PSA) were purchased from Thermo
Fisher. Other qPCR Taqman sets were designed using the Universal Probe
Library System (Roche) and are available in Supplementary Data 4.

Bader D.A., Hartig S.M., Putluri V., Foley C., Hamilton M.P., Smith E.A., Saha P.K., Panigrahi A., Walker C., Zong L., Martini-Stoica H., Chen R., Rajapakshe K., Coarfa C., Sreekumar A., Mitsiades N., Bankson J.A., Ittmann M.M., O’Malley B.W., Putluri N, & McGuire S.E. (2018). Mitochondrial pyruvate import is a metabolic vulnerability in androgen receptor-driven prostate cancer. Nature metabolism, 1(1), 70-85.

+ Open protocol

+ Expand

Quantifying RNA Expression in Cell Lines

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

For validation of Multi-miR shRNAs in NIH/3T3 cells, total RNA from cells was extracted using The RNeasy Kit and RNase-Free DNase Set (Qiagen) according to the manufacturer’s instructions. For Multi-miR mediated inhibition of tumor suppressor genes in the mouse liver, total RNA from liver tumors was extracted using the AllPrep DNA/RNA Micro Kit and RNase-Free DNase Set (Qiagen) according to the manufacturer’s instructions. cDNA synthesis was prepared from 1 μg total RNA using Taqman reverse transcription kit (Applied Biosystems, #N808-0234) with random hexamers. For comparison of LT3GEPIR and pCF806 efficacy, RNA isolation and cDNA generation was performed using the Cells-to-CT kit (Invitrogen, #4402954), per the manufacturer’s instructions. qRT-PCR analyses were carried out in technical triplicate using SYBR green (Applied Biosystems) and specific primers (Table S1). Measurements were carried out using the ViiA seven system (Life Technologies) or a QuantStudio five Real-Time PCR machine (Thermo Fisher Scientific). The mRNA expression levels were normalized to the levels of mouse Actb mRNA, or human GUSB mRNA, and quantified using the comparative C_T method.

Amen A.M., Loughran R.M., Huang C.H., Lew R.J., Ravi A., Guan Y., Schatoff E.M., Dow L.E., Emerling B.M, & Fellmann C. (2022). Endogenous spacing enables co-processing of microRNAs and efficient combinatorial RNAi. Cell Reports Methods, 2(7), 100239.

+ Open protocol

+ Expand

Spheroid Co-Culture: Hepatocyte and Endothelial RNA Isolation

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

After 9 days of culturing, co-cultured spheroids composed of hepatocytes and
Endothelial cells were retrieved from the PDMS microwells and from the 3D ECM
scaffold after collagenase treatment for 12 h (Collagenase type 1,
1 mg ml⁻¹). RNA was purified using the
RNeasy kit (Qiagen, CA, USA) and cDNA was generated using reverse transcriptase
(TAKARA, Japan) according to the manufacturer's instructions. The primer
sequences are listed in the Supplementary Table 1.

Jeong G.S., No D.Y., Lee J., Yoon J., Chung S, & Lee S.H. (2016). Viscoelastic lithography for fabricating self-organizing soft micro-honeycomb structures with ultra-high aspect ratios. Nature Communications, 7, 11269.

+ Open protocol

+ Expand

qPCR Analysis of Gene Expression

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

+ Open protocol

+ Expand

Overexpression of Cd274 in B16-F0 cells

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

Total RNA was isolated from the LN8-1205BL cell line using the
RNEasy kit (Qiagen) and converted to cDNA with the iScript cDNA synthesis
kit (BioRad). Two rounds of PCR were performed to amplify the
Cd274 CDS and add a Kozak sequence and restriction
sites. The primers were as follows: PCR1 forward:
5’-GCCACCATGAGGATATTTGCTGGCATTATATTC-3’, PCR1 reverse:
5’-CGGGAATTCTTACGTCTCCTCGAATTGTGTATC-3’, PCR2 forward:
5’-ATTACTAGTGCCGCCACCATGAGGATATTTGC-3’, PCR2 reverse:
5’-TCTAGACCCGGGAATTCTTACGTCTCCTCG-3’. The product was then
cloned into the pLVX-EF1a-IRES-Hygro plasmid described above by restriction
digest. Lentiviral particle production and viral transduction were performed
as described above. Transduced cells were selected in Hygromycin B followed
by sorting, as described above. Control cells were transduced with a pLVX
vector lacking the Cd274 gene. B16-F0-cntrl or
–Cd274 cells were transplanted into wild-type
female mice as described above. Mice were euthanized 26 days following cell
implantation and LN metastasis incidence was quantified.

Reticker-Flynn N.E., Zhang W., Belk J.A., Basto P.A., Escalante N.K., Pilarowski G.O., Bejnood A., Martins M.M., Kenkel J.A., Linde I.L., Bagchi S., Yuan R., Chang S., Spitzer M.H., Carmi Y., Cheng J., Tolentino L.L., Choi O., Wu N., Kong C., Gentles A.J., Sunwoo J.B., Satpathy A.T., Plevritis S.K, & Engleman E.G. (2022). Lymph node colonization induces tumor-immune tolerance to promote distant metastasis. Cell, 185(11), 1924-1942.e23.

+ Open protocol

+ Expand

Tissue-Specific RNA Extraction and qPCR Analysis

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

Total RNA from muscle tissue models was extracted using FastPrep FP120
(Qbiogene, USA) homogenization for frozen samples in combination with the
RNeasy kit (Qiagen, Hilden, Germany). Total RNA from tendon models was
extracted using Freezer/Mill 6870 (SPEXSamplePrep; Metuchen, NJ, USA) and
TRIZOL (ThermoFisher) extraction. RNA expressions were determined by
quantitative PCR (qPCR) using the High Capacity cDNA kit (Lithuana),
Universal PCR Master mix, and corresponding TaqMan Assays, all from Applied
Biosystems (USA). Muscle gene expressions were normalized using
the geomean of 18S RNA, GAPDH, TBP, and β2M housekeeping gene expressions.
Tenomodulin expression was normalized using Eif4a2 housekeeping gene
expression. Three tissue models were analyzed per time point, with the
exception of day 4 muscle proliferation models (n = 2).
Mean and standard error of mean (SEM) were calculated. qPCR analysis has
been repeated three times for muscle and tendon tissue models in independent
experiments to verify reproducibility of differentiation and tissue
engineering.

Laternser S., Keller H., Leupin O., Rausch M., Graf-Hausner U, & Rimann M. (2018). A Novel Microplate 3D Bioprinting Platform for the Engineering of Muscle and Tendon Tissues. Slas Technology, 23(6), 599-613.

+ 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!