RNAsimple Total RNA Kit

RNA isolation / purification Tissue - Rat Placenta

Experiment
RNA isolation / purification Tissue - Rat Placenta
Product
RNAsimple Total RNA Kit from Tiangen
Manufacturer
Tiangen

Protocol tips

Upstream tips
- To wipe off RNase, the glassware can be roasted at 150℃ for 4 hours, while plastic can be dipped in 0.5 M NaOH for 10min, washed by RNase-Free ddH2O thoroughly, and sterilized

Publication protocol

Total RNA of placenta from each group was extracted using Trizol and RNA simple Total RNA Kit (Tiangen Biotech, Beijing, China). RNA was quantified by UV absorption measure and 1 µg was reverse transcribed to cDNA. The reverse transcription (RT) was carried out using PrimeScript RT reagent Kit (TaKaRa, Dalian, China) for 15 min at 37°C, 5 sec at 85°C in a 20 µl reaction volume. Real-time PCR was performed using SYBR Premix Ex Taq (TaKaRa) according to the manufacturer’s instruction. Primer sequences were ordered from Sangon (Shanghai, China) (Table 1). The real-time PCR cycling condition for GAPDH, CYP1A1, VEGF-A, VEGF-B, PIGF, Flt-1, fetal liver kinase-1 (Flk-1), soluble Flt-1 (sFlt-1), HIF-1α, Endoglin and IFN-γ is 30 sec at 95°C for incubation, 40 cycles of 15 sec at 95°C and 20 sec at 60°C. To confirm the amplification specificity, the PCR products were subjected to a melting curve analysis. Levels of mRNA expression were analyzed using 2−△△CT method and gene expression was normalized to GAPDH.

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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 - Rat Placenta using RNAsimple Total RNA Kit from Tiangen.

Paper title
ITE and TCDD Differentially Regulate the Vascular Remodeling of Rat Placenta via the Activation of AhR
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Manufacturer protocol

Download the product protocol from Tiangen for RNAsimple Total RNA Kit below.

Download PDF Download manufacturer protocol

Videos

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