RNeasy Plus Mini Kit

RNA isolation / purification Tissue - Mouse Hypothalamus

Experiment
RNA isolation / purification Tissue - Mouse Hypothalamus
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

For gene expression analyses, mRNA was obtained from the hypothalamus tissue of adult mice and GT1-7 cells using RNeasy Plus Mini Kits (Qiagen, Chatsworth, CA, US). We used 1000 ng total mRNA as the template for cDNA synthesis primed with random hexamers (Bio-Rad, Richmond, CA, US). Then, the reaction mixture was diluted at 1:2, and 2 μl of the diluted template was used per 20 μl of the real-time quantitative PCR assay using the Bio-Rad SsoAdvanced™ Universal SYBR (Bio-Rad 172–5274, Mississauga, ON, Canada). The Bio-Rad CFX Connect Real-Time system and the Bio-Rad CFX Manager Software (version 2.0) were used to collect the PCR data. Results are presented as linearized values normalized against β-actin. The primers (Table S2) were synthesized by the Beijing Genomic Institute (BGI).



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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Mouse Hypothalamus using RNeasy Plus Mini Kit from Qiagen.

Paper title
Nasal delivery of nerve growth factor rescue hypogonadism by up-regulating GnRH and testosterone in aging male mice
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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