NucleoSpin® RNA

RNA isolation / purification Tissue - Rat Jejunum

Experiment
RNA isolation / purification Tissue - Rat Jejunum
Product
NucleoSpin® RNA from Macherey Nagel
Manufacturer
Macherey Nagel

Protocol tips

Upstream tips
- Aliquot rDNase and store at -20 °C.
Protocol tips
Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Publication protocol

RNA extraction was performed on jejunum and colon samples as per manufacturer’s instructions (NucleoSpin RNA Isolation Kit, Macherey-Nagel). Briefly, tissue samples were homogenized in 250 μl and 500 μl of TRIzol® Reagent (Invitrogen), respectively. Samples were centrifuged for 15 min at 4 °C and the upper aqueous layer removed. Following a sequence of filtration steps, RNA-binding conditions were adjusted and DNA digestion performed. A series of washing steps was performed before highly pure mRNA was eluted in RNase-free water. Once eluted, RNA was stored at −80 °C.

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Discussion

Discussion

1 year 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

2 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

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Manufacturer protocol

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