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Index / Biolin / TRIsure / Product Detail : BIO-38032 TRIsure
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Sep
25th

#BIO-38032 TRIsure

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  Price : 186   EUR
211   USD
144   GBP
781   Zloty
24906   JPY
1435   NOK
1538   SEK
210   CHF

Product name : TRIsure

Catalog number : BIO-38032

Quantity: 100ml

Availability: Yes

Supplier name : Biolin

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More Details about

 

Features

□  □ Quick isolation of high-quality RNA

□  □ Ready-to-use solution for a wide variety cells and tissues

□  □ Cost-effective

□  □ Convenient 1hr protocol

□  □ Performs well with large or small amounts of tissue or cells Applications

□  □ Isolation of high quality RNA from diverse biological material, including animal and plant tissues rich in polysaccharides and proteoglycans

□  □ Purified RNA is ideal for any downstream application such as RT-PCR or in vitro translation

Description

TRIsure is a ready-to-use reagent for the isolation of total RNA from cells and tissues. TRIsure maintains the integrity of the extracted RNA, while disrupting

cells and subsequently dissolving cell components. The reagent combines a blend of phenol and other components.

The biological sample is homogenized or lysed in TRIsure and then separated into organic and aqueous phases. The RNA remains in the aqueous phase

and is subsequently recovered by precipitation with isopropyl alcohol.

The isolated RNA is suitable for any downstream application such as RT-PCR, hybridization assays, or in vitro

translation. 1ml of TRIsure is sufficient to

isolate Total RNA from 1 x 107 cells or 100mg of tissue.

The expected yield of RNA from 1mg of tissue is:

□  □ 2-5 ^g from mouse kidney

□  □ 5-10^g from mouse liver

The expected yield of RNA from 1 x 106 cultured cells is:

□  □ 8-15^g from epithelial cells

□ □ 20-25^g from cell fibroblasts

Protocol for the Isolation of RNA using TRIsure Reagents Required:

□  □ Chloroform

□  □ Isopropyl alcohol (chilled)

□  □ 75% Ethanol (in DEPC-treated water)

               □ DEPC-treated water 1. Homogenization

Tissue:

Homogenize tissue samples in 1 ml of TRIsure per 50-100mg of tissue. For small quantities of tissue (1-10mg), add 800^l of TRIsure. For samples of fat tissue, a layer of fat may accumulate at the top, which should be removed.

Plant tissue

Following homogenization, insoluble material is removed by centrifugation at 12,000 x g for 10 minutes at 2-8°C. Transfer the cleared homogenate to a fresh tube.

Cells Grown on Monolayer:

Lyse cells directly in a culture dish or flask by adding 1ml of TRIsure per 10cm2 growth area, pipette the cell lysate several times to ensure sufficient cell disruption.

Cells Grown in Suspension:

Pellet cells at 200 x g for 5 minutes at room temperature. Lyse cells with 1 ml of TRIsure per 5 x 106 cells and pass the lysate several times through a pipette tip. For small quantities of cells (102 -106), lyse cells in 800^l of TRIsure.

Note: At this stage, samples can be stored for at least one month at -60 to -70°C. Description

TRIsure is a ready-to-use reagent for the isolation of total RNA from cells and tissues. TRIsure maintains the integrity of the extracted RNA, while disrupting

cells and subsequently dissolving cell components. The reagent combines a blend of phenol and other components.

The biological sample is homogenized or lysed in TRIsure and then separated into organic and aqueous phases. The RNA remains in the aqueous phase

and is subsequently recovered by precipitation with isopropyl alcohol.

The isolated RNA is suitable for any downstream application such as RT-PCR, hybridization assays, or in vitro

translation. 1ml of TRIsure is sufficient to

isolate Total RNA from 1 x 107 cells or 100mg of tissue.

The expected yield of RNA from 1mg of tissue is:

□  □ 2-5 ^g from mouse kidney

□  □ 5-10^g from mouse liver

The expected yield of RNA from 1 x 106 cultured cells is:

□  □ 8-15^g from epithelial cells

□  □ 20-25 ^g from cell fibroblasts

2.  Phase Separation

Incubate samples for 5 minutes at room temperature. Add 0.2 ml of chloroform per 1 ml of TRIsure used. Cap tubes securely and shake vigorously by hand for 15 seconds.

Incubate samples for 2-3 minutes at room temperature. Centrifuge samples at 12,000 x g for 15 minutes (or 2600 x g for 20-30 minutes) at 2-8°C. The sample will separate into a pale green, phenol-chloroform phase, an interphase, and a colorless upper aqueous phase that contains the RNA.

3.  RNA Precipitation

Transfer the aqueous phase very carefully, without disturbing the interphase to another tube. Precipitate the RNA by mixing with cold isopropyl alcohol. Use 0.5 ml of isopropyl alcohol per 1 ml of TRIsure used. Incubate samples for 10 minutes at room temperature then centrifuge at 12,000 x g for 10 minutes (or 2600 x g for 20-30 minutes) at 2-8°C.

Note: For small quantities of cells, RNase-free Glycogen Co-precipitant (BIO-37077) can be added to the aqueous phase before addition of isopropyl alcohol to aid RNA precipitation. Add 5-10^g of Glycogen per 800^l of TRIsure.

4.  RNA Wash

Remove the supernatant. Wash the pellet once with 75% ethanol, adding at least 1 ml of ethanol per 1 ml of TRIsure used. Vortex samples and centrifuge at 7500 x g for 5 minutes at 2-8°C.

Note: At this stage, samples can be stored for one week at 2-8°C, or 12 months at -5 to -20°C.

5. Re-dissolving the RNA

Air-dry the pellet and dissolve in RNase free water (BIO-37080) or DEPCtreated Water (BIO-38030) by pipetting the solution up and down. Incubate for 10 minutes at 55-60°C if necessary. Store RNA at -70°C.

 

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