Methanobacterium Transduction Agar

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanobacterium Transduction Agar

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

Composition per liter:

Agar solution..........................................................................500.0mL
Basal salts solution.................................................................100.0mL
Na 2 CO 3 solution.......................................................................24.0mL
Titanium (III) citrate solution ..................................................16.0mL
Trace elements solution ...........................................................10.0mL
L -Cysteine/Na 2 S solution.........................................................10.0mL
Agar Solution:
Composition per 500.0mL:
Agar ............................................................................................15.0g
Preparation of Agar Solution:  Add agar to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Gently heat and
bring to boiling. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 60°C while sparging with 80% N 2 + 20%
CO 2 .
Basal Salts Solution:
Composition per liter:
KH 2 PO 4 .......................................................................................68.0g
NH 4 Cl .........................................................................................21.2g
Resazurin .................................................................................10.0mg
Preparation of Basal Salts Solution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 .
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................9.55g
MgCl 2 ·7H 2 O................................................................................4.06g
FeCl 2 ·7H 2 O.................................................................................0.98g
NaWO 4 ·6H 2 O............................................................................0.264g
NiCl 2 ·6H 2 O...............................................................................0.118g
Na 2 Se 2 O 3 .......................................................................................0.1g
NaMoO 4 ·2H 2 O..........................................................................0.024g
CoCl 2 ·6H 2 O ..............................................................................0.023g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 7.0 with
KOH. Add remaining components one at a time. Add distilled/deion-
ized water to 1.0L. Adjust pH to 7.0. Sparge with 100% N 2 .
Na 2 CO 3 Solution:
Composition per liter:
Na 2 CO 3 .......................................................................................10.6g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N 2 + 20% CO 2 .
L -Cysteine/Na 2 S Solution:
Composition per 50.0mL:
L -Cysteine·HCl..............................................................................5.0g
Na 2 S·9H 2 O....................................................................................5.0g
Preparation of  L -Cysteine/Na 2 S Solution:  Gently heat and

bring 50.0mL of distilled/deionized water to boiling. Add  L -

cysteine·HCl and Na 2 S·9H 2 O. Mix thoroughly. Cool while sparging
with 100% N 2 . Bring volume to 50.0mL with distilled/deionized water.
Sparge with 100% N 2 . Adjust pH to 10.0 with 3N NaOH.
Titanium (III) Citrate Solution:
Composition per 55.0mL:
Sodium citrate·9H 2 O.....................................................................2.9g
Titanium (III) chloride................................................................0.75g
Preparation of Titanium (III) Citrate Solution:  Add compo-
nents to distilled/deionized water and bring volume to 55.0mL. Mix
thoroughly. Filter sterilize. Sparge with 100% N 2 .
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except agar solution and titanium cit-
rate solution, to distilled/deionized water and bring volume to
484.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 50°–55°C. Aseptically
and anaerobically add 55.0mL of sterile titanium citrate solution to
500.0mL of sterile agar solution. Add the agar/titanium/citrate solution
to the rest of the medium. Mix thoroughly. Aseptically and anaerobi-
cally, under an atmosphere of 80% H 2 + 20% CO 2 , distribute into ster-
ile Petri dishes, sterile tubes, or sterile bottles.
Use: For the cultivation of Methanobacterium species for transduc-
tion.