RNeasy Plus Mini Kit

RNA isolation / purification Cells - immortalized Cal-27

Experiment
RNA isolation / purification Cells - immortalized Cal-27
Product
RNeasy Plus Mini Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Downstream tips
- Include DNAse treatment for 15-20min.

- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C.

- Use water to elute the RNA that is warmed to ~60`C.

Publication protocol

HNSCC cells were seeded in 60 mm dishes (2 × 105 cells/dish) and supernatants were collected and saved for secreted protein analysis. Cells were washed twice with PBS before isolating total RNA using RNeasy Plus mini kit (QIAGEN) as per manufacturer's protocol. 500 ng of isolated RNA was then reverse transcribed to cDNA using iScript cDNA synthesis kit (Bio‐Rad) and a thermocycler under the following conditions: 5 min at 25° C, 30 min at 42° C, and 5 min at 85° C. The resultant cDNA samples were used to perform quantitative (i.e. real time) PCR analysis in a 96‐well optical plate with each well containing 6 μL of cDNA, 7.5 μL of SyBr Green Universal SuperMix (Bio‐Rad), and 1.5 μL oligonucleotide primers (forward and reverse; 4 μM) for a total reaction volume of 15 μL) on an ABI PRISM Sequence Detection System (model 7000, Applied Bio systems) with the following protocol: 95 °C for 15 s (denaturing) and 60 °C for 60 s (annealing) repeated for 40 cycles. Oligonucleotide primers were obtained from Integrated DNA Technologies (IDT; Coralville, IA) and described in Supplementary Table 1. Relative gene expression was determined by the comparative 2ˆ (−ΔΔCT) method. Briefly, cycle threshold (CT) values of all genes were normalized to that of GAPDH for each sample (in duplicate) and then fold changes were determined by comparing the normalized CT values of erlotinib‐sensitive with erlotinib‐resistant HNSCC cells. Each assay was performed in triplicate (i.e. on three separate occasions) and the results were presented as mean ± standard error of the mean.

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

Reviews

RNeasy Plus Mini Kit from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

2 years ago

Author: Switzerland

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized Cal-27 using RNeasy Plus Mini Kit from Qiagen.

Paper title
Upregulated interleukin‐6 expression contributes to erlotinib resistance in head and neck squamous cell carcinoma
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Plus Mini Kit below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Cells - immortalized Cal-27 using RNeasy Plus Mini Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Become shareholder Discussions About us Contact Privacy Terms