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File name:YTSV厌氧液体培养基制备
Category:Protocols
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Published:2013-12-11 03:44:08
Update:2013-12-11 03:56:04
Summary:

一、试剂

1、1000× Cysteine-HCl

      称取12.5 g Cysteine-HCl· XH2O粉末溶解在少量纯水中,定容到50 ml。
                     
2、1000× Na2S

      称取12.5 g Na2S· 9 H2O溶解在少量纯水中,定容到50 ml。                      

3、Trace element solution A (g/L):

CuSO4· 6H2O                                0.01

ZnSO4· 7H2O                                  0.1

CoCl2· 6H2O                                 5 mg 
 
MgCl2· 4H2O                                  0.2

Na2MoO4· 2H2O                             0.1

KBr                                                 0.05

KI                                                   0.05

H3BO3                                             0.1

LiCl                                                0.05

Al2(SO4)3                                      0.05

NiCl2· 6H2O                                 0.01

4、Trace element solution B (g/L):

VoSO4· H2O                                 0.05

H2WO                                        0.05

Na2SeO3                                       0.05

NiCl2· 6H2O                                0.05

SrCl2· 6H2O                                0.05

BaCl2                                           0.05

5、(NH4)2SO4 and KH2PO4溶液(g/L):

(NH4)2SO                                  43

KH2PO4                                      3.6

6、FeEDTA溶液(g/L): 

FeSO4· 7H2O                            1.54

Na2EDTA                                  2.06

7、人工海水 (g/L):

NaCl                                          19.6
 
Na2SO4                                       3.3
 
KCl                                             0.5
 
KBr                                           0.05
 
H3BO3                                      0.02
 
MgCl2· 6H2O                             8.8

8、YTSV培养基 (pH 6.5)

Trace element solution A                       10ml

Trace element solution B                          1ml

(NH4)2SO4 and KH2PO4溶液               10ml

FeEDTA溶液                                           2ml

Yeast Extract                                              1 g

Tryptone                                                    1 g

Element sulfur                                             5g

人工海水                                           补至1 L

二、实验步骤

1、按照YTSV培养基配方添加各成分,用约800 ml人工海水充分溶解。

2、按1 ml/L加入1000× Cysteine-HCl母液和1000× Na2S母液到培养基中,再加入刃天青溶液作氧指示剂,

      混匀,调节pH至所需值,将培养基定容至1L。

3、元素硫分装成0.5g/瓶,培养基按100ml/瓶分装,向血清瓶中充入纯氮约3 min,盖上橡胶塞和铝盖,

      用专用起子将铝盖密封在瓶口。

4、121℃灭菌20min,放置过夜,如果培养基颜色不变红则可以使用。

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