Description

The kit adopts magnetic beads and a unique buffer system, to isolate and purify total RNA with high quality from cell/tissue. The uniquely embedded magnetic beads have a strong affinity for nucleic acid under certain conditions. When the conditions are changed, the magnetic beads can release the absorbed nucleic acid to rapidly separate and purify it.

It is compatible with automated nucleic acid extractors. Through absorption, transfer and release of magnetic beads by the special magnetic bar, magnetic beads and nucleic acid can be transferred to improve the degree of automation. The whole process is safe and convenient, and the extracted RNA has high purity. If high throughput automated extraction is required, Biofargo can deliver integrated solutions.

RNA purified by this kit is suitable for downstream experiments including RT-PCR, Real Time RT-PCR, chip analysis, Northern Blot, Dot Blot, PolyA screening, in vitro translation, RNase protection analysis and molecular cloning.

Kit Contents

Group	Component	GDP771 (96 preps)
GDP771H	RNAstore Reagent	12 ml
	Buffer TSA	70 ml
	Proteinase K	2 × 1 ml
	Buffer ICBP	35 ml
	MagAttract Suspension BE (100 mg/ml)	2 × 1 ml
	Buffer RDC	90 ml
	Buffer RW1A	200 ml
	Buffer RW4	90 ml
	Buffer RW	20 ml
	RNase-Free ddH₂O	30 ml
GRT431	RNase-Free DNase I (1500 U)	1 pcs
GRT431	RNase-Free ddH₂O	1 ml

Note: GRT431, GDP771H are shipped and stored separately.

Features

Simple and fast: Ultra-pure total RNA can be obtained quickly.
Safe and non-toxic: No toxic reagents such as phenol/chloroform.
High purity: The RNA obtained has high purity and can be directly used for chip detection, high-throughput sequencing and other experiments.

Reagents need to be prepared by Customer

Isopropanol (supplementary protocol: samples preserved in Trizol), anhydrous ethanol, homogenization equipment (mortar, homogenizer, etc.), liquid nitrogen, gloves, masks, RNase-Free centrifuge tubes, magnetic racks, or nucleic acid extractor, for other brands of extractor to be integrated, please contact Biofargo.

Recommended dosage for different types of samples

Note: Please refer to the recommended ranges in the table below. Do not exceed 100 mg of tissue, otherwise it may lead to RNA yield and quality decrease.

Category	Sample Type	Optimal Extraction Amount
Tissue	Spleen	5 mg
	Liver	10 mg
	Kidney	15 mg
	Intestine	15 mg
	Gland	15 mg
	Heart	25 mg
	Lung	25 mg
	Brain	25 mg
	Fat	50 mg
	Skin	100 mg
	Muscle	100 mg
	Tail	0.3 cm (rat), 0.6 cm (mouse)
Cell	Non-stop protocol	10⁵ – 10⁷ cells
Cell	Low yield, pause protocol recommended	Less than 10⁵ cells
Insects	Mollusc	25 mg
Insects	Crustacea	50 mg
Fungus	Yeast	Not more than 5 × 10⁷ cells
	Mould	25 mg
	Mushrooms	75 mg

Storage

All components of the kit GDP771H can be stored in dry conditions at room temperature (15-30°C) for 15 months. RNase-Free DNase I and RNase-Free ddH2O can be stored at 2-8°C for 15 months.

Important Notes

Replace the gloves with new ones frequently. This is because skin often carries bacteria that can lead to RNase contamination.
Avoid cross-contamination by using RNase-free plastics and tips.
Pay attention to optimal sample storage and pretreatment conditions to avoid causing degradation of the extracted RNA

Preparation of DNase I storage solution

Dissolve DNase I dry powder (1500 U) in 1 ml RNase-Free ddH2O, mix gently and follow the protocol to add 10 μl to each sample for freshly use.

Attention: If long-term storage needed, dissolve DNase I dry powder (1500 U) in 550 μl RNase-Free ddH2O to make stock solution, mix gently, dispense and store at -30~-15°C (can be stored for 9 months). Add 5 μl stock solution to each sample. Store the DNase I stock solution after thawing from -30~-15°C at 2-8°C (can be stored for 6 weeks) and do not freeze it again.

Magnetic Hi-Tissue/Cell Total RNA Kit