RP Medium

[所属分类:培养基配方] [发布时间:2021-8-6] [发布人:网站管理员2] [阅读次数:] [返回]

RP Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Solution 1...............................................................................960.0mL
Solution 2.................................................................................40.0mL
Solution 1:
RPMI 1640 solution...............................................................900.0mL
HEPES solution .......................................................................60.0mL
RPMI 1640 Medium:
Composition per liter:
Inorganic salt solution............................................................400.0mL
Other component solution......................................................400.0mL
Amino acid solution...............................................................100.0mL
Vitamin solution.....................................................................100.0mL
Inorganic Salt Solution:
Composition per 400.0mL:
NaCl..............................................................................................6.0g
NaH 2 PO 4 ·H 2 O...............................................................................0.8g
KCl................................................................................................0.4g
Ca(NO 3 ) 2 ·4H 2 O.............................................................................0.1g
MgSO 4 (anhydrous)...............................................................48.84mg
Preparation of Inorganic Salt Solution: Add components to dis-
tilled/deionized water and bring volume to 400.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Other Component Solution:
Composition per 400.0mL:
D -Glucose......................................................................................2.0g
Phenol Red.................................................................................5.0mg
Glutathione, reduced..................................................................1.0mg
Preparation of Other Component Solution:  Add components
to distilled/deionized water and bring volume to 400.0mL. Mix thor-
oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Amino Acid Solution:
Composition per 100.0mL:
L -Glutamine...........................................................................300.0mg
L -Arginine..............................................................................200.0mg
L -Cysteine·2HCl.......................................................................65.0mg
L -Asparagine (free base)..........................................................50.0mg
L -Isoleucine..............................................................................50.0mg
L -Leucine.................................................................................50.0mg
L -Lysine·HCl............................................................................40.0mg
L -Serine....................................................................................30.0mg
L -Tyrosine·2Na·2H 2 O..............................................................29.0mg
L -Aspartic acid.........................................................................20.0mg
L -Glutamic acid .......................................................................20.0mg
L -Hydroxyproline ....................................................................20.0mg
L -Proline ..................................................................................20.0mg
L -Threonine..............................................................................20.0mg
L -Valine....................................................................................20.0mg
L -Histidine (free base) .............................................................15.0mg
L -Methionine............................................................................15.0mg
L -Phenylalanine .......................................................................15.0mg
Glycine.....................................................................................10.0mg
L -Tryptophan .............................................................................5.0mg
Preparation of Amino Acid Solution: Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Filter sterilize.
Vitamin Solution:
Composition per 100.0mL:
i-Inositol...................................................................................35.0mg
Folic acid ...................................................................................1.0mg
Niacinamide...............................................................................1.0mg
Para-aminobenzoic acid.............................................................1.0mg
Pyridoxal·HCl............................................................................1.0mg
Pyridoxine·HCl..........................................................................1.0mg
Thiamine·HCl............................................................................1.0mg
Choline chloride.........................................................................3.0mg
D -Ca pantothenate....................................................................0.25mg
Biotin.........................................................................................0.2mg
Riboflavin..................................................................................0.2mg
Vitamin B 12 ............................................................................0.005mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of RPMI 1640 Medium: Aseptically combine 400.0mL
of sterile inorganic salt solution, 400.0mL of sterile other component
solution, 100.0mL of sterile amino acid solution, and 100.0mL of ster-
ile vitamin solution.
HEPES Solution:
Composition per 60.0mL:
HEPES........................................................................................5.94g
Preparation of HEPES Solution: Add HEPES to distilled/deion-
ized water and bring volume to 60.0mL. Mix thoroughly. Filter steril-
ize.
Preparation of Solution 1:  Aseptically combine 960.0mL of sterile
RPMI-1640 solution and 60.0mL sterile HEPES solution.
Solution 2:
Composition per 100.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of Solution 2:  Add NaHCO 3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: Aseptically combine 960.0mL of sterile
solution 1 with 40.0mL of sterile solution 2. Aseptically distribute into
sterile tubes or flasks.
Use: For the cultivation of Balamuthia mandrillaris and Plasmodium
falciparum.