CTAB Extraction Buffer (RNase free) - BioReagent,Suitable for molecular biology

Cat. No.: C1518317

GRADE & PURITY Suitable for molecular biology, BioReagent

Synonyms

CTAB Lysis Buffer (RNase-free) | CTAB Isolation Buffer (RNase-free) | CTAB RNA Extraction Buffer | RNase-free CTAB Buffer

Storage

Protected from light,Room temperature

Shipped In

Normal

Application

RNA Extraction

Overview

  Common methods for extracting genomic DNA from plant tissues include cesium chloride centrifugation and CTAB extraction. The CTAB extraction method is a classic technique for plant nucleic acid extraction. This product is RNase-free and primarily used for total RNA extraction from plant tissues. The nucleic acids obtained can meet the requirements of most molecular biology experiments.

  The active component of CTAB Extraction Buffer (RNase-free) is CTAB (cetyltrimethylammonium bromide). It is treated to be RNase-free, and 2-ME should be added before use to enhance the reagent's effectiveness and stability. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or other applications.

C1518317	Component	500 mL	Storage
C1518317A	CTAB Extraction Buffer (RNase-free)	500 mL	RT.
C1518317B	2-ME	10 mL	RT. Store in the dark.

Materials to Prepare

• Laboratory equipment: Liquid nitrogen, mortar or homogenizer, centrifuge tubes, incubator or water bath, centrifuge
• Reagents: DEPC-treated water, chloroform/isoamyl alcohol (24:1), 75% ethanol (prepared with DEPC-treated water)

Procedure (For Reference Only)

1. Take 5 mL or an appropriate amount of CTAB Extraction Buffer and mix it with 2-ME at a ratio of 50:1 (CTAB Extraction Buffer:2-ME). Place the mixture in a 15 mL or other suitable centrifuge tube and preheat at 60°C. If necessary, add 1–5 μg/mL of DNase to remove DNA from the sample.

2. Weigh 1.0–1.5 g or an appropriate amount of fresh plant tissue or leaves. Pre-cool the mortar or homogenizer with liquid nitrogen or dry ice, and grind the fresh plant tissue or leaves into a fine powder. Transfer the frozen tissue into a centrifuge tube.

3. Add 4–5 mL/g of preheated CTAB Extraction Buffer to the powdered tissue, mix thoroughly, and incubate at 65°C for 15–60 minutes, mixing occasionally.

4. Add an equal volume of chloroform/isoamyl alcohol (24:1), mix thoroughly by inverting, and centrifuge at 8000 g for 5–10 minutes. Recover the upper aqueous phase (supernatant), which contains the desired DNA/RNA.

5. Transfer the supernatant to a new centrifuge tube, add 1/2 to 2/3 volumes of pre-chilled isopropanol, mix gently, and let it stand at room temperature to allow nucleic acids to settle at the bottom of the tube. If no precipitate is observed, let it stand at room temperature for several hours or overnight.

6. Centrifuge at 2000 g for 2 minutes, and carefully discard the supernatant.

7. Add 75% ethanol to the loose DNA/RNA pellet, let it stand at room temperature for 20 minutes, and centrifuge at 4000 g for 10 minutes. Carefully discard the supernatant.

8. Air-dry the RNA naturally and dissolve it in an appropriate amount of deionized water or TE buffer. If necessary, add 1–5 μg/mL of DNase I to remove residual DNA. Store at -20°C.

Notes

1. If the usage amount is small each time, the reagent can be aliquoted before use.

2. For your safety and health, please wear a lab coat and disposable gloves during operation.

3. Use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.

Specifications

Synonyms

CTAB Lysis Buffer (RNase-free) | CTAB Isolation Buffer (RNase-free) | CTAB RNA Extraction Buffer | RNase-free CTAB Buffer

Specifications & Purity

BioReagent, Suitable for molecular biology

Stability And Storage

Store at room temperature long term (12 months). Store in the dark.

Storage

Protected from light, Room temperature

Shipped In

Normal

Grade

BioReagent, Suitable for molecular biology

Chemical and Physical Properties

• Sensitivity: Light-sensitive