Luận văn : Áp dụng qui trình nuôi chín noãn in vitro trên heo và chó part 6 pps

46
Điều chỉnh pH = 7,4 bằng NaOH 1N hoặc NaOH 5N.
(f) Môi trƣờng khi nuôi
FCS 200 μl
Pyruvic acid 200 μl
hMG 200 μl
TCM 199 1400 μl
2. Trên chó
(a) Thành phần môi trƣờng
TCM199 môi trường căn bản
HEPES 25mM/l
ECS 10%
Gentamycin 50 μg/ml
NaHCO
3
2,2 mg/ml
Pyruvic acid 22μg/ml
Estradiol 1 μg/ml
FSH 0,5 μg/ml
hCG 0,03 UI/ml
(b) Pha môi trƣờng căn bản TCM199
Nước cất 100ml
TCM199 980 mg
NaHCO
3
220 mg
Gentamycin 5 mg
Hepes 5,97g
Trữ ở 4
o

C.
(c) Pha NaHCO
3

TCM199 10ml
NaHCO
3
0,22g
Trữ ở -20
o
C.
(d) Pha pyruvic acid
TCM199 10 ml
Pyruvic acid 0,022g
Trữ ở -20
o
C.

47
(e) Pha estradiol, FSH, hCG bằng môi trường căn bản TCM199. Trữ ở -20
o
C.
(f) Môi trƣờng nuôi trứng chín
Pyruvic acid 50 μl
Estradiol 50 μl
FSH 50 μl
hMG 50 μl
TCM199 4300 μl
3. Môi trƣờng PBS – PVA
Nước cất 100 ml

NaCl 800 mg
KCl 20 mg
Na
2
HPO
4
.12H
2
O 290 mg
KH
2
PO
4
20 mg
Polyvinyl alcohol (PVA) 100 mg
PVA được pha riêng với nước cất, gia nhiệt khoảng 70
o
C đến khi PVA tan hết trong
nước. Sau đó được pha chung với môi trường PBS – PVA.

48
PHỤ LỤC 2. BẢNG TRONG STAGRAPHIC


Analysis of Variance for HEO.LOAIA - Type I Sums of Squares


Source of variation Sum of Squares d.f. Mean square F-ratio Sig.
level


MAIN EFFECTS
A:HEO.PPHAP .0591267 1 .0591267 .479 .4971
B:HEO.KTHUOC 1.8879429 2 .9439714 7.652 .0007

RESIDUAL 21.341147 173 .1233592


TOTAL (CORRECTED) 23.288217 176


0 missing values have been excluded.
All F-ratios are based on the residual mean square error.

Table of Least Squares Means for HEO.LOAIA


95%
Confidence
Level Count Average Stnd. Error for mean


GRAND MEAN 177 .4825215 .0478110 .3881326.5769104
A:HEO.PPHAP
1 84 .4960188 .0556274 .3861985.6058390
2 93 .4690242 .0536471 .3631135.5749349
B:HEO.KTHUOC
1 90 .4164220 .0370971 .3431846.4896594
2 80 .6235000 .0392682 .5459763.7010237
3 7 .4076425 .1328045 .1454584.6698265

49
Analysis of Variance for HEO.LOAIB - Type I Sums of Squares


Source of variation Sum of Squares d.f. Mean square F-ratio Sig.
level


MAIN EFFECTS
A:HEO.PPHAP .0341841 1 .0341841 .351 .5609
B:HEO.KTHUOC 1.1146264 2 .5573132 5.716 .0039

RESIDUAL 16.868994 173 .0975086


TOTAL (CORRECTED) 18.017805 176


0 missing values have been excluded.
All F-ratios are based on the residual mean square error.

Table of Least Squares Means for HEO.LOAIB


95%
Confidence
Level Count Average Stnd. Error for mean


GRAND MEAN 177 .3639194 .0425073 .2800011.4478377
A:HEO.PPHAP
1 84 .3535265 .0494567 .2558887.4511643
2 93 .3743123 .0476960 .2801503.4684743
B:HEO.KTHUOC
1 90 .4377429 .0329819 .3726297.5028560
2 80 .2755000 .0349121 .2065761.3444239
3 7 .3785153 .1180724 .1454155.6116151

50
Analysis of Variance for HEO.loaiC - Type I Sums of Squares


Source of variation Sum of Squares d.f. Mean square F-ratio Sig.
level


MAIN EFFECTS
A:HEO.PPHAP .0099814 1 .0099814 .146 .7066
B:HEO.KTHUOC .2376642 2 .1188321 1.742 .1782

RESIDUAL 11.800240 173 .0682095


TOTAL (CORRECTED) 12.047886 176


0 missing values have been excluded.
All F-ratios are based on the residual mean square error.

Table of Least Squares Means for HEO.loaiC


95%
Confidence
Level Count Average Stnd. Error for mean


GRAND MEAN 177 .1619216 .0355520 .0917345.2321087
A:HEO.PPHAP
1 84 .1711807 .0413643 .0895189.2528425
2 93 .1526625 .0398918 .0739078.2314172
B:HEO.KTHUOC
1 90 .1691564 .0275852 .1146974.2236153
2 80 .1010000 .0291996 .0433538.1586462
3 7 .2156084 .0987528 .0206497.4105672

51

Analysis of Variance for CHO5.A - Type I Sums of Squares


Source of variation Sum of Squares d.f. Mean square F-ratio Sig.
level


MAIN EFFECTS
A:CHO5.buongtrung .0001369 1 .0001369 .002 .9652
B:CHO5.kichthuoc .0086363 1 .0086363 .124 .7295

RESIDUAL 3.5399402 51 .0694106

TOTAL (CORRECTED) 3.5487134 53


0 missing values have been excluded.
All F-ratios are based on the residual mean square error.













Table of Least Squares Means for CHO5.A


95%
Confidence
Level Count Average Stnd. Error for mean


GRAND MEAN 54 .7903141 .0377690 .7144723.8661558
A:CHO5.buongtrung
1 26 .7855320 .0557147 .6736546.8974095
2 28 .7950961 .0498658 .6949635.8952287

B:CHO5.kichthuoc
1 35 .8039689 .0448452 .7139177.8940200
2 19 .7766593 .0619596 .6522418.9010767
















52
Analysis of Variance for CHO5.B - Type I Sums of Squares


Source of variation Sum of Squares d.f. Mean square F-ratio Sig.
level


MAIN EFFECTS
A:CHO5.buongtrung .0001369 1 .0001369 .002 .9652
B:CHO5.kichthuoc .0086363 1 .0086363 .124 .7295

RESIDUAL 3.5399402 51 .0694106


TOTAL (CORRECTED) 3.5487134 53


0 missing values have been excluded.
All F-ratios are based on the residual mean square error.









Table of Least Squares Means for CHO5.B


95%
Confidence
Level Count Average Stnd. Error for mean


GRAND MEAN 54 .2096859 .0377690 .1338442.2855277
A:CHO5.buongtrung
1 26 .2144680 .0557147 .1025905.3263454
2 28 .2049039 .0498658 .1047713.3050365

B:CHO5.kichthuoc
1 35 .1960311 .0448452 .1059800.2860823
2 19 .2233407 .0619596 .0989233.3477582