小鼠生殖工程学技术——13试剂的组成
试剂的组成包括 1M DMSO，磷酸二铵213，0.25M蔗糖，mHTF和KSOM
回到小鼠生殖工程学技术指南目录
回到小鼠生殖工程冻存及培养试剂
1M DMSO
|
试剂成分 |
mg/100ml |
供应商 |
目录编号 |
|
NaCl |
555 |
Sigma |
S 5886 |
|
KCl |
18.5 |
Sigma |
P 5405 |
|
KH2PO4 |
4.75 |
Sigma |
P 5655 |
|
MgSO4 7H2O |
4.95 |
Sigma |
M 7774 |
|
CaCl2 2H2O |
25 |
Sigma |
C 7902 |
|
NaHCO3 |
210 |
Sigma |
S 5761 |
|
Glucose |
3.6 |
Sigma |
G 6152 |
|
Na-Pyruvate |
2.2 |
Sigma |
P 4562 |
|
DL-Lactic Acid, sodium salt |
0.174ml |
Sigma |
L 1375 |
|
10mM EDTA |
100μl |
Sigma |
E-6635 |
|
Streptomycin |
5 |
Sigma |
S 9137 |
|
Penicillin |
6.3 |
Sigma |
P 7794 |
|
0.5% phenol red |
0.1ml |
Sigma |
P 0290 |
|
L-Glutamine |
14.6 |
Sigma |
G 8540 |
|
MEM Essential Amino Acids |
1ml |
Sigma |
11130-051 |
|
MEM Non-essential AA |
0.5ml |
Sigma |
M 7145 |
|
BSA |
100 |
Sigma |
A-4378 |
参考文献
【1】 Nakao K., Nakagata N., and Katsuki M. 1997. Simple and effcient procedure for cryopreservation
of mouse embryosby simple vitrification. Exp. Anim. 46: 231-234.
DAP213
2M DMSO，1M乙酰胺，含3M丙二醇的PB1
PB1
|
试剂成分 |
mg/100ml |
供应商 |
目录编号 |
|
NaCl |
800.0 |
Sigma |
S5886 |
|
KCl |
20.0 |
Sigma |
P5405 |
|
CaCl2 |
12.0 |
Sigma |
C5670 |
|
KH2PO4 |
20.0/td> |
Sigma |
P5655 |
|
MgCl2 6H2O |
10.0 |
Sigma |
M2393 |
|
Na2HPO4 |
115.0 |
Sigma |
S5136 |
|
Na-Pyruvate |
3.6 |
Sigma |
P4562 |
|
Glucose |
100.0 |
Sigma |
G6152 |
|
Penicillin |
7.5 |
P4687 |
L 1375 |
|
Streptomycin |
5.0 |
Sigma |
S1277 |
|
BSA |
300.0 |
Sigma |
A4378 |
溶液A
|
试剂名称 |
mL (最终体积10mL) |
供应商 |
目录编号 |
|
PB1 |
2.3088 |
– |
– |
|
DMSO |
3.1252 |
Sigma |
D2650 |
|
Propylene glycol (PG) |
4.556 |
Sigma |
P1009 |
溶液B
|
试剂名称 |
mg/10mL in PB1 |
供应商 |
目录编号 |
|
Acetamide (AA) |
1181.4 |
Sigma |
A0500 |
DAP213 : 首先准备A溶液和B溶液，并让各自完全溶解。将等量的A、B溶液混合一起形成DAP213.
注意：加入DMSO时，溶液可能变得混浊。
参考文献
【1】 Nakao K., Nakagata N., and Katsuki M. 1997. Simple and effcient procedure for cryopreservation of
mouse embryos by simple vitrification. Exp. Anim. 46: 231-234.
0.25M 蔗糖
PB1
|
试剂成分 |
mg/100ml |
供应商 |
目录编号 |
|
NaCl |
800.0 |
Sigma |
S 5886 |
|
KCl |
20.0 |
Sigma |
P-5405 |
|
CaCl2 |
12.0 |
Sigma |
C 5670 |
|
KH2PO4 |
20.0 |
Sigma |
P5655 |
|
MgCl2 6H2O |
10.0 |
Sigma |
M 2393 |
|
Na2HPO4 |
3.6 |
Sigma |
S 5136 |
|
Na-Pyruvate |
3.6 |
Sigma |
P-4562 |
|
Glucose |
100.0 |
Sigma |
G-6152 |
|
Penicillin |
7.5 |
Sigma |
P-4687 |
|
Streptomycin |
5.0 |
Sigma |
S-1277 |
|
BSA |
300.0 |
Sigma |
A-4378 |
0.25M Sucrose in PB1
|
试剂成分 |
mg/20ml in PB1 |
供应商 |
目录编号 |
|
Sucrose |
1711.5 |
Sigma |
S-1888 |
参考文献
【1】 Nakao K., Nakagata N., and Katsuki M. 1997. Simple and effcient procedure for cryopreservation of
mouse embryos by simple vitrification. Exp. Anim. 46: 231-234.
mHTF
|
试剂成分 |
mg/100ml |
供应商 |
目录编号 |
|
NaCl |
593.8 |
Sigma |
S 5886 |
|
KCl |
35.0 |
Sigma |
P5405 |
|
MgSO4 7H2O |
4.9 |
Sigma |
M7774 |
|
KH2PO4 |
5.4 |
Sigma |
P5655 |
|
CaCl2 |
57.0 |
Sigma |
C5670 |
|
NaHCO3 |
210.0 |
Sigma |
S5761 |
|
Glucose |
50.0 |
Sigma |
G6152 |
|
Na-lactate (mL)* |
0.34mL |
Sigma |
L7900 |
|
Na-Pyruvate |
3.7 |
Sigma |
P4562 |
|
Penicillin G |
7.5 |
Sigma |
P4687 |
|
Streptomycin |
5.0 |
Sigma |
S1277 |
|
BSA(Albumin, Bovine serum, Fraction V, Fatty Acid-Free) |
400 |
MERCK/CALBIOCHEM |
126575 |
|
0.5% phenol red (mL) |
0.04mL |
Sigma |
P0290 |
* 含量： 70%
参考文献
【1】 Kito S., Hayao T., Noguchi-Kawasaki Y., Ohta Y., Hideki U., and Tateno S. 2004. Improved in vitro
fertilization and development by use of modified human tubal fluid and applicability of pronucleate
embryos for cryopreservation by rapid freezing in inbred mice. Comp. Med. 54(5): 564-570.
KSOM
|
试剂成分 |
mg/100ml |
供应商 |
目录编号 |
|
NaCl |
555 |
Sigma |
S 5886 |
|
KCl |
18.5 |
Sigma |
P 5405 |
|
KH2PO4 |
4.75 |
Sigma |
P 5655 |
|
MgSO4 7H2O |
4.95 |
Sigma |
M 7774 |
|
CaCl2 2H2O |
25 |
Sigma |
C 7902 |
|
NaHCO3 |
210 |
Sigma |
S 5761 |
|
Glucose |
3.6 |
Sigma |
G 6152 |
|
Na-Pyruvate |
2.2 |
Sigma |
P 4562 |
|
DL-Lactic Acid, sodium salt |
0.174ml |
Sigma |
L 1375 |
|
10mM EDTA |
100μl |
Sigma |
E-6635 |
|
Streptomycin |
5 |
Sigma |
S 9137 |
|
Penicillin |
6.3 |
Sigma |
P 7794 |
|
0.5% phenol red |
0.1ml |
Sigma |
P 0290 |
|
L-Glutamine |
14.6 |
Sigma |
G 8540 |
|
MEM Essential Amino Acids |
1ml |
Gibco |
11130-051 |
|
MEM Non-essential AA |
0.5ml |
Sigma |
M 7145 |
|
BSA |
100 |
Sigma |
A-4378 |
参考文献
【1】 Lawitts J.A., and Biggers J.D. 1993. Culture of preimplantation embryos. Methods Enzymol. 225:153-164.