Inzol Reagent(TRIzol)(IC-5299)

Background

Inzol Reagent is a product that can extract RNA from animal and plant samples. The sample can be fully lysed in Inzol Reagent. During the homogenization or lysis of the sample, it can maintain the integrity of the RNA, while lysing the cells and dissolving the cell contents. Inzol Reagent has a strong broad spectrum and can be applied to the extraction of total RNA from various samples. The extraction process is convenient and fast, and the entire operation can be completed within one hour. This reagent can be used for small samples (50-100 mg tissue, 1x106 cells) and also suitable for large samples (¡Ý1g tissue, >107 cells). It is applicable to humans, animals, plant tissues, and bacteria, and can process a large number of different samples at the same time, and the entire extraction process can be completed within one hour. The isolated total RNA protein and DNA are extremely low in contamination, and can be used for Northern Blot, reverse transcription, polyA screening, in vitro translation, RNase protection analysis, and gene cloning.

Self-Provided Reagents

Chloroform, isopropanol, 70% ethanol (DEPC water configuration), Rnase Free H2O.

Size

100ML

Inzol usage instructions

Sample pretreatment

Adherent cells

1. Pour out the culture solution from every 10 cm2 of the cultured cells and wash them with PBS once to remove as much excess solution as possible.

2. Add 1 ml of Inzol to distribute the lysate evenly on the cell surface, and then use a pipette to blow the cells off. Transfer the cell lysate to a 1.5ml EP tube.

Suspension cells, yeast, bacteria

1. Pour the suspension cultured cells together with the culture solution into a centrifuge tube, centrifuge at 8,000 rpm for 2 min, discard the supernatant, and add 50¦Ìl of sterile water to resuspend the cells until there is no obvious precipitation.

2. Add 1ml Inzol.

 Animal and Plant tissue

1. ransfer the sample to a mortar precooled with liquid nitrogen, grind the tissue with a pestle, and continuously add liquid nitrogen in the meantime until it is ground into a powder.

2. Add the ground tissue to a 1.5 ml EP tube containing 1 ml Inzol.

Total RNA Extraction

1. After adding Inzol, immediately turn it upside down with wrist force until the cells and tissue powder are evenly dispersed without lumps. Leave it at room temperature for 5 minutes to completely separate the nucleic acid-protein complexes.

2. Add 200 ¦Ìl chloroform, shake vigorously with the wrist for 15 seconds, and leave it at room temperature for 2 minutes .

3. Centrifuge at 12,000 rpm for 15 minutes, and transfer colorless  supernatant to a new 1.5EP tube.

4. Add 600 ¦Ìl of isopropanol to the supernatant, turn it upside down several times vigorously with the wrist,  Centrifuge at 12,000 rpm for 15 min.

5.  carefully discard the supernatant, Add 1ml of 70% ethanol to each tube of the pellet, turn it upside down several times, and centrifuge at 7,500 rpm for 5 min. Carefully discard the supernatant and save the bottom RNA pellet.

6. Repeat step 5 and wash again.

7. Pour off the washing solution, centrifuge again for a short time for 10 seconds, absorb

the remaining washing solution with a 10 ¦Ìl tip, and place it at room temperature to evaporate the ethanol (~20min).

8. Add 20-100¦Ìl TE Buffer or RNase Free H2O to each tube to dissolve total RNA.

 Troubleshooting

Storage

Inzol Reagent should be stored at 4¡æ

Note: For your safety and health, please wear experimental clothes and disposable gloves.This product is for scientific research only.Inzol Reagentis for research use only. It is not approved for human or animal use, or for application in invitrodiagnostic procedures.