Regeneration Dextrose Medium, w/o Agar | RD medium | 250g

Regeneration Dextrose Medium, w/o Sorbitol & Agar | RD medium | 250g

For the version with 500g Sorbitol (in bottle B), please request a quote and order via support@gmexpression.com

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 and so on.

• 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^ 186g
  
• 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

^ Customer prepared or in Bottle B of RD Medium with Sorbitol version(to be purchased separately).

Preparation

Dissolve 186 g of sorbitol (prepared separately by the customer or in Bottle B of RD Medium with Sorbitol version) in 900 mL of deionized water. Add 20g of agar (or agarose) if preparing solid media. autoclave at 121°C for 20 min, then place in a 45°C water bath to maintain temperature.

Dissolve 33.43 g of RD medium in 100 mL of deionized water by heating, sterilize by 0.22 µm filtration and preheat to 45°C in a water bath.

Mix the above two solutions. If culturing his4 strain, add sterilized 10 mL of 0.4% histidine solution (prepared separately by customer).