Regeneration Dextrose Medium, w/o Agar | RD medium | 250g
- Product Code: GMNB-RD01
- Availability: In Stock
Tags: RD medium
Regeneration Dextrose Medium, w/o Sorbitol & Agar | RD medium | 250g
If need Sorbitol, please select it in the purchase options. We also provide Pichia Spheroplast Preparation Kit with Zymolyase (Lyticase from Arthrobacter Luteus), please contact us via support@gmexpression.com for more details.
Cautions
This media is prepared in small batches and will be prepared after the purchase, taking around 5 to 10 business days for handling.
Use
Regeneration Dextrose Medium (RD medium, RDB Agar) is a liquid medium used for the regeneration of protoplasts of Pichia Pastoris competent cells; it can also be used to screen nutrient-deficient (transformed with vector contains HIS4 gene) Pichia Pastoris transformants. Suitable his4 strains include GS115, KM71, SMD1168, etc.
- Add 2% Agar to RD medium can make RDB agar.
- Add 1% agarose to RD medium can make RD top agar.
- Add L-histidine to RD medium can make RDH medium.
- Add both L-histidine and 2% Agar to RD medium can make RDBH agar.
- Add L-histidine and 1% agarose to RD medium can make RDH top agar.
Components, per litre
|
Sorbitol[1] |
186 g |
|
Glucose |
20 g |
|
YNB |
13.4 g |
|
D-Biotin |
0.4 mg |
|
L-Glutamic acid |
0.005 g |
|
L-Methionine |
0.005 g |
|
L-Lysine |
0.005 g |
|
L-Leucine |
0.005 g |
|
L-Isoleucine |
0.005 g |
[1]. Customer prepared or in Bottle B of RD Medium with Sorbitol version(to be purchased separately).
Preparation
- Sorbitol Solution:
Dissolve 186 g of sorbitol, which needs to be prepared separately by the customer or contained in Bottle B of RD Medium with Sorbitol version, in 900 mL[2] of deionised or Milli-Q (MQ) water. If preparing solid media, add 20 g of agar or agarose. Autoclave the solution at 121°C for 20 minutes, then maintain the temperature at 45°C using a water bath.
- RD Medium Solution:
Dissolve 33.43 g of RD Medium in 100 mL[2] of deionised or MQ water by heating. Sterilise the solution using a 0.22 µm filtration method, then preheat to 45°C in a water bath.
- Mix the above two solutions. If culturing his4 strain, add sterilised 10 mL of 0.4% histidine solution (prepared separately by the end user).
[2]. Users may adjust these volumes. For instance, dissolving sorbitol in 800 mL of water while filtering 200 mL of RD Medium solution to mitigate the low filtration rate of concentrated solutions
