RNAiso Plus

RNAiso Plus is a total RNA extraction reagent that can easily and quickly isolate RNA from plant and animal tissues or cultured cells. After adding RNAiso Plus solution to disrupt tissues or cells, add chloroform to the disrupted solution, mix well, and centrifuge to separate into three layers. At this time, RNA is in the upper transparent water layer, DNA is in the semi-solid middle layer, and finally, the lower organic solvent layer that is red * contains proteins, polysaccharides, fatty acids, cell debris, and a small amount of DNA. Here, only the upper transparent water layer can be collected and total RNA can be recovered by isopropanol precipitation.

Using this product, the entire total RNA extraction process can be completed in about 1 hour, and the separated total RNA can be used for RT-PCR ** , Northern blot analysis, mRNA separation, and in vitro translation reactions.

* Since the organic solvent layer in this product is red, it is easy to distinguish the boundary, making it more convenient to use.

** When using purified RNA samples for RT-PCR, treatment with Recombinant DNase I (RNase-free) (Code 2270A) before use is recommended because even a small amount of gDNA contamination can significantly affect the results .

Specifications

• Effective total RNA extraction from plant and animal tissues and cultured cells

• RNA extraction using AGPC (Acid Guanidinium-Phenol-Chloroform)

• Optimal for cases where large amounts of samples are required or large amounts of total RNA are required.

Detail

RNAiso Plus (Code 9108)*	100 ml
RNAiso Plus (Code 9109)*	200 ml

* Since it contains protein denaturants, be careful not to let it come into contact with skin or clothing. If it gets into your eyes or on your skin, immediately wash with plenty of running water and seek medical advice.

Preservation

4℃ (shade preservation)

Reagents required other than this product

- Chloroform

- Isopropanol

- 75% ethanol [ prepared using DEPC-treated H2O] -

RNase-free water

RNAiso Plus Usage Guidelines

Type and quantity of samples	RNAiso Plus Usage (ml)
10 cm^2 of adherent cells 5×10^6 ~1×10^7 suspended cells 100 μl of white blood cells	1~2 1 2
50~100 mg of tissue sample - Tissues from which RNA extraction is relatively easy - Tissues from which high-purity RNA extraction is difficult (liver, spleen, bone, and cartilage *1, etc.)	1 2
15~30 mg of plant material *2 (contains small amounts of polysaccharides and phenols)	1
2~5x10^7 yeast cells *3	1

*1 For bone and cartilage tissue samples, we recommend using High-Salt Solution for Precipitation (Plant) (Code 9193).

*2 For plant samples containing a lot of polysaccharides, we recommend using the pretreatment reagent Fruit-mate™ for RNA Purification (Code 9192).

*3 For yeast samples, we recommend using the pretreatment reagent Yeast Processing Reagent (for total RNA preparation) (Code 9089).

Average total RNA extraction amount of RNAiso Plus

Group	Sample Quantity	Total RNA Extraction Amount
Between the mouse	1 g	About 4,000~5,000 µg
Mouse kidney	1 g	About 3,000 µg
Mouse skeletal muscle	1 g	About 1,500 µg
Mouse brain	1 g	About 1,500 µg
HL60 cultured cells	1×10^7 pieces	About 100 µg
Tobacco leaves	1 g	About 1,000 µg
White blood cells	1×10^7 pieces	About 20~40 µg
Whole blood *	1 ml	15~20 µg
Carp skeletal muscle	1 g	About 50 µg

Use 1 ml of RNAiso Plus per 100 µl of whole blood.

RNAiso Plus for various samples

Sample	Thesis Title	Paper Information
Drosophila	Curcumin-supplemented diets increase superoxide dismutase activity and mean lifespan in Drosophila	Age (Dordr) 2013 August; 35(4): 1133?1142.
Fruit moth	Identification of Putative Olfactory Genes from the Oriental Fruit Moth Grapholita molesta via an Antennal Transcriptome Analysis	PLoS One. 2015; 10(11): e0142193.
Small intestines	Human Lipocalin-Type Prostaglandin D Synthase-Based Drug Delivery System for Poorly Water-Soluble Anti-Cancer Drug SN-38	PLoS One. 2015; 10(11): e0142206.
Bone tissue	CD44 deficiency inhibits unloadinginduced cortical bone loss through downregulation of osteoclast activity	Sci Rep. 2015; 5: 16124.
Penicillium	Whole transcriptome analysis of Penicillium digitatum strains treated with prochloraz reveals their drug-resistant mechanisms	BMC Genomics. 2015; 16: 855.
Drosophila melanogaster	Molecular Mechanisms for High Hydrostatic Pressure-Induced Wing Mutagenesis in Drosophila melanogaster	Sci Rep. 2015; 5: 14965.
Bombyx larvae	Molecular Mechanisms for High Hydrostatic Pressure-Induced Wing Mutagenesis in Drosophila melanogaster	Sci Rep. 2015; 5: 13839.
Fishing Spiders	A Comparative Analysis of the Venom Gland Transcriptomes of the Fishing Spiders Dolomedes mizhoanus and Dolomedes sulfurous	PLoS One. 2015; 10(10): e0139908.
Seedling	RNAi-based functional elucidation of PtrPRP, a gene encoding a hybrid proline rich protein, in cold tolerance of Poncirus trifoliata	Front Plant Sci. 2015; 6:808.
Cucumber fruit	QTL mapping of cucumber fruit flesh thickness by SLAF-seq	Sci Rep. 2015; 5: 15829.

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