Plant protoplast preparation and transformation kit
â— Product composition:
Component number | name | specification | Storage | transport |
RTU4052-01 | Solution I (2×)-enzyme dissolution solution | 100 ml | -20 ° C | RT |
RTU4052-02 | Solution II-W5 Solution | 200 ml | -20 ° C | RT |
RTU4052-03 | Solution III-MMG Solution | 25 ml | -20 ° C | RT |
RTU4052-04 | Solution IV-conversion solution | 2.5 ml | -20 ° C | RT |
RTU4052-05 | Solution V-WI Solution | 25 ml | -20 ° C | RT |
BME | reducing agent | 1 ml | RT | RT |
BSA-02 | 50mg/ml BSA solution | 5 ml | -20 ° C | RT |
Instruction manual | One |
â— Product introduction:
Plant protoplasts refer to vital cells with vitality that are removed from the plasma membrane by removing all cell walls. It has cell life characteristics and versatility, is an important source of cell clonal variation and mutant screening, and is also an ideal material for plant genetic engineering and an ideal material for genetic improvement. Enzymatic separation of protoplasts is a common technique. The principle is that plant cell walls are mainly composed of cellulose, hemicellulose and pectin, so cellulase, hemicellulase, eductase and pectinase can be used to degrade cell walls. The components are removed from the cell wall to obtain protoplasts. Many methods can induce protoplast fusion, and the fusion method that is now widely used is the polyethylene glycol (PEG) method. As a polymer compound, polyethylene glycol (PEG) can produce a transient impact on protoplasts at a suitable concentration. Protoplasts quickly shrink and adhere, and then washed with high calcium and high pH method to fuse protoplasts. carry out.
The kit contains all the reagents required for the preparation and transformation of plant protoplasts other than enzymes, and can be used 20 times according to the use of 10 ml of enzyme solution at a time.
â— Storage and expiration date : stored according to temperature, valid for one year.
â— Instructions for use:
Materials to be prepared (not supplied with the kit):
Cellulase R-10; isolation enzyme R-10; flathead scorpion; 70 μm cell filter; disposable blade; 50 ml centrifuge tube; 1.5 ml centrifuge tube;
First, the separation of protoplasts:
Ready-to-use enzyme solution
10ml formula | 20 ml formula | |
Solution I (2×) | 5 ml | 10 ml |
Cellulase R-10 | 0.15 grams | 0.3 grams |
Separation enzyme R-10 | 0.04 grams | 0.08 grams |
reducing agent | 5 μl | 10 μl |
50mg/ml BSA | 0.2 ml | 0.4 ml |
Sterilized water | Make up to 10 ml | Make up to 20 ml |
2. After enzymatic hydrolysis, add 10 ml of Solution II and mix gently.
3. Filter the solution from step 2 with a 70 μm mesh screen to remove undigested leaves and collect the filtrate in a 50 ml centrifuge tube.
4. Centrifuge 100 g of the filtrate for 2 minutes at room temperature to remove the supernatant.
5. Resuspend the protoplasts with 1 ml of solution II and ice for 30 minutes (the protoplasts will naturally precipitate at the bottom).
6. Carefully remove the supernatant solution, do not touch the protoplast precipitate, and resuspend the pellet in 1 ml of solution III to be the protoplast solution.
Second, the original plastid transformation:
1. Take 10 μl of plasmid DNA (concentration 1-2 μg/μl) in a 1.5 ml centrifuge tube.
2. Add 100 μl of protoplast solution and mix gently.
3. Add an equal volume of 110μl solution IV-transformation solution, gently mix thoroughly, leave it at room temperature for 15 minutes, mix gently and intermittently.
4. Add 2 times the volume, ie 440 μl of Solution II, gently and thoroughly mix to terminate the conversion process.
5. Centrifuge at 100g for 2 minutes at room temperature and remove the supernatant.
6. Resuspend the pellet in 1 ml of solution V.
3. Protoplast culture and collection:
1. The protoplast solution is incubated at room temperature (20-25 ° C) for 2-16 hours.
Note: The incubation time is determined according to the experimental requirements. RNA analysis was incubated for 2-6 hours; enzymatic activity analysis and protein labeling experiments were incubated for 2-16 hours.
2. The protoplasts were collected by centrifugation at 100 g for 2 minutes at normal temperature to remove most of the supernatant.
3. The protoplast pellet was resuspended in the remaining solution and stored at -80 °C.
Medical Cold Patch
Patch for diarrhea
[Name] Medical Cold Patch
[Package Dimension] 5cm 4pieces/box
The pain relief patch is composed of three layers, namely, backing lining, middle gel and protective film. It is free from pharmacological, immunological or metabolic ingredients.
[Scope of Application] For cold physiotherapy, closed soft tissue only.
[Indications]
The patches give a fast relief for diarrhea.
[How To Use a Patch]
Please follow the Schematic Diagram. One piece, one time.
The curing effect of each piece can last for 6-8 hours.
[Attention]
Do not apply the patch on the problematic skin, such as wounds, eczema, dermatitis,or in the eyes. People allergic to herbs and the pregnant are advised not to use the medication. If swelling or irritation occurs, please stop using and if any of these effects persist or worsen.notify your doctor or pharmacist promptly. Children using the patch must be supervised by adults.
[Storage Conditions]
Store below 30c in a dry place away from heat and direct sunlight.
Patch For Diarrhea,Medicated Patches For Arthiritis,Plaster For Diarrhea,Pad For Diarrhea
Shandong XiJieYiTong International Trade Co.,Ltd. , https://www.sdxjmedical.com