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

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Bên dưới lớp màng là noãn nguyên thủy. Khi nang noãn chín, tế bào nang bao quanh noãn phân chia thành nhiều tầng tế bào hạt. Nang noãn càng phát triển thì tế bào nang phân hủy tạo ra một xoang chứa dịch. Khi trở nên chín, nang noãn được bao bọc bởi một lớp màng mỏng.

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

  1. 46 Điều chỉnh pH = 7,4 bằng NaOH 1N hoặc NaOH 5N. (f) Môi trƣờng khi nuôi 200 μl FCS 200 μl Pyruvic acid 200 μl hMG 1400 μl TCM 199 2. Trên chó (a) Thành phần môi trƣờng môi trường căn bản TCM199 HEPES 25mM/l ECS 10% 50 μg/ml Gentamycin NaHCO3 2,2 mg/ml 22μg/ml Pyruvic acid 1 μg/ml Estradiol 0,5 μg/ml FSH hCG 0,03 UI/ml (b) Pha môi trƣờng căn bản TCM199 Nước cất 100ml TCM199 980 mg NaHCO3 220 mg Gentamycin 5 mg Hepes 5,97g Trữ ở 4oC. (c) Pha NaHCO3 TCM199 10ml NaHCO3 0,22g Trữ ở -20oC. (d) Pha pyruvic acid TCM199 10 ml Pyruvic acid 0,022g Trữ ở -20oC.
  2. 47 (e) Pha estradiol, FSH, hCG bằng môi trường căn bản TCM199. Trữ ở -20oC. (f) Môi trƣờng nuôi trứng chín 50 μl Pyruvic acid 50 μl Estradiol 50 μl FSH 50 μl hMG 4300 μl TCM199 3. Môi trƣờng PBS – PVA Nước cất 100 ml NaCl 800 mg KCl 20 mg Na2HPO4.12H2O 290 mg KH2PO4 20 mg Polyvinyl alcohol (PVA) 100 mg PVA được pha riêng với nước cất, gia nhiệt khoảng 70oC đến khi PVA tan hết trong nước. Sau đó được pha chung với môi trường PBS – PVA.
  3. 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 --------------------------------------------------------------------------- ---
  4. 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 --------------------------------------------------------------------------- ---
  5. 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 --------------------------------------------------------------------------- ---
  6. 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 --------------------------------------------------------------------------- ---
  7. 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 --------------------------------------------------------------------------- ---
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