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 . .3 861 985 .60 58390 2 93 . 469 0242 .05 364 71 . 363 1135.5749349 B :HEO. KTHUOC 1 90 .4 164 220 .0370971 .34318 46. 48 965 94 2 80 .62 35000 .039 268 2 .5459 763 .7010237 3 7 .40 764 25 .1328045 .1454584 .66 98 265 . .0895189.2528425 2 93 .15 266 25 .0398918 .0739078.2314172 B :HEO. KTHUOC 1 90 . 169 1 564 .0275852 .11 469 74.22 361 53 2 80 .1010000 .02919 96 .0433538.15 864 62 3 7 .21 560 84 .0987528 .02 064 97.410 567 2 . MEAN 54 .7903141 .037 769 0 .7144723. 866 1558 A:CHO5.buongtrung 1 26 .7855320 .0557147 .67 365 46. 8974095 2 28 .7950 961 .049 865 8 .69 4 963 5.8952287 B:CHO5.kichthuoc 1 35 .803 968 9 .0448452 .7139177.8940200