Total RNA Isolation Kit (Blood/Cultured Cell/Fungus)

Catalog numberPDC04-0100

CategoriesColumn Based

Size100 Reactions

Sample4 rxns ($31.2)

$528 Inquire

Note: PureDireX / Purification Kits / Extraction Kit / RNA

Description

The Total RNA Isolation Kit provides a fast, simple, and cost-effective method for the isolation of total RNA from the whole blood, mammalian cells, and bacterial cells. Detergents and chaotropic salt are used to lyse cells and inactivate RNase. The specialized high-salt buffering system allows RNA species bases to bind to the glass fiber matrix of the spin column while contaminants pass through the column. Impurities are efficiently washed away, and the pure RNA is eluted with the REL Buffer without phenol extraction or alcohol precipitation. The RNA purified with the Total RNA Isolation Kit is suitable for a variety of routine applications, including the RT-PCR, cDNA Synthesis, Northern Blotting, Differential display, Primer Extension and mRNA selection. The entire procedure can be completed within 25-40 minutes.

Sample:

Up to 300 μl of whole bloodUp to 10⁷ mammalian cells

Up to 10⁹ bacterial cellsUp to 10⁸ fungus cells

Format: Reagent and spin column
Yield: Up to 30 μg
Operation time: 25-40 minutes
Elution volume: 50-200 ul

Kit Contents

Contents	PDC04-0100	PDC04-0100S (Sample)
Buffer RL	110 ml	4 ml
Buffer RA	45 ml	2 ml
Buffer RO	25 ml	1 ml
Buffer W1	45 ml	2 ml
Buffer W2 (Add ethanol)	15 ml (60 ml)	300 µl × 2 (1.5 ml × 2)
Buffer RE	10 ml	1 ml
Column DR	100 pcs	4 pcs
Collection Tubes	100 pcs	4 pcs

Features

Delivers high-quality total RNA with the fast procedure.
Ready-to-use RNA for high performance in any downstream application.
Consistent RNA yield from the starting material with a small amount.

Protocol

Fresh Blood/Cultured Mammalian Cells
Bacterial Cells/Fungus Cells

Total RNA Isolation Kit (Blood/Cultured Cell/Fungus)

Troubleshooting

Refer to the table below to troubleshoot problems that you may encounter when purifying total RNA.

Problem	Cause	Solution
Degraded RNA/ low integrity	RNases contaminant	Clean everything, use barrier tips, wear gloves and a lab coat, and use RNase-free enzymes, EX: RNase inhibitor.
Low yields of RNA	Incomplete lysis and homogenization	Use the appropriate method for the lysate preparation based on the amount of the starting materials immersed in the Buffer RA to achieve the optimal lysis.
Low yields of RNA	Incorrect elution conditions	Add 50 μl of the RE Buffer to the center of each Column DR, let it stand for 2 minutes, and centrifuge at 14,000 x g for 2 minutes.
Inhibition of downstream enzymatic reactions	Presence of ethanol in the purified RNA	Repeat the wash step: Centrifuge at 14,000 x g again for 2 minutes to remove the residual W2 Buffer.

Caution

During the operation, always wear the latex or vinyl gloves while handling reagents and RNA samples to prevent the RNase contamination.
Add 60 ml of the ethanol (96~100%) to the Buffer W2, and shake before use (see bottle label for volume).
Check Buffers before use for salt precipitation. Re-dissolve any precipitate by warming up to 37°C.
The Buffers RA and W1 contain irritants., so please wear gloves when handling these buffers.
Research Use Only. Not intended for any animal or human therapeutic or diagnostic uses.

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