Sea Urchin Vitelline Envelope Removal

Devin Morris

Maurice Fernandez

Milton Diaz

Question

Do the two different methods used in VE removal (Alpha-Amylase & DTT) yield the same data in 10% and 12% gels,

or does one yield more accurate results than the others?

Removal of Vetelline Envelope

Vitelline Envelope

- Outer membrane of

female egg that plays

a role in fertilization

α-Amylase

- Enzymes used to break

up starches

Dithiothreitol - DTT- Reduces disulfide

bonds in proteins and

enzymes.

Manual

- Manually remove the Vitelline Envelope

Gels Measured

10% Acrylamide gel

Concentration

12% Acrylamide gel

Concentration

Lanes

- Measured each band length

- Adjusted brightness and contrast on lanes to make better measurements of bands

10% Manual 12% Manual

5.35.25.15.04.94.84.74.64.5

4

3

2

1

0

Log MW

RM

S = 0.221238 R-Sq = 96.7 % R-Sq(adj) = 95.9 %

RM = 19.8255 - 3.66571 Log MW

Regression Plot

Linear

4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3

0.5

1.5

2.5

3.5

Log MW

RM

RM = 579.976 - 335.455 Log MW

+ 65.2819 Log MW**2 - 4.26682 Log MW**3

S = 0.0328031 R-Sq = 100.0 % R-Sq(adj) = 99.9 %

Regression Plot

Quadratic

4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3

0.5

1.5

2.5

3.5

Log MW

RM

RM = 579.976 - 335.455 Log MW

+ 65.2819 Log MW**2 - 4.26682 Log MW**3

S = 0.0328031 R-Sq = 100.0 % R-Sq(adj) = 99.9 %

Regression Plot

Cubic

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1676 0.1595 0.1644 0.1653 0.1603 0.1644 0.1603 0.1644 0.1622116250.0000 5.0654 cm band 2 0.3243 0.3189 0.3208 0.3198 0.3179 0.3181 0.3179 0.3208 0.318997400.0000 4.9886 cm band 3 0.4000 0.3919 0.3962 0.3902 0.3886 0.3908 0.3913 0.3935 0.391966200.0000 4.8209 cm band 4 0.5270 0.5216 0.5310 0.5257 0.5245 0.5229 0.5217 0.5202 0.527045000.0000 4.6532 cm band 5 0.7514 0.7432 0.7466 0.7480 0.7418 0.7439 0.4728 0.7493 0.748631000.0000 4.4914 cm band 6 0.9919 0.9865 0.9919 0.9864 0.9864 0.9838 0.9891 0.9811 0.9811

SSError 0.0127 0.0126 0.0123 0.0127 0.0125 0.0129 0.0792 0.0126 0.0116DegFreedom 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000StdDev'n 0.0564 0.0560 0.0554 0.0564 0.0560 0.0567 0.1407 0.0561 0.0538R-sqAdj 0.9630 0.9634 0.9643 0.9630 0.9635 0.9626 0.7692 0.9634 0.9663R-squared 0.9704 0.9707 0.9714 0.9704 0.9708 0.9701 0.8153 0.9707 0.9730

10% Linear Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1676 0.1595 0.1644 0.1653 0.1603 0.1644 0.1603 0.1644 0.1622116250.0000 5.0654 cm band 2 0.3243 0.3189 0.3208 0.3198 0.3179 0.3181 0.3179 0.3208 0.318997400.0000 4.9886 cm band 3 0.4000 0.3919 0.3962 0.3902 0.3886 0.3908 0.3913 0.3935 0.391966200.0000 4.8209 cm band 4 0.5270 0.5216 0.5310 0.5257 0.5245 0.5229 0.5217 0.5202 0.527045000.0000 4.6532 cm band 5 0.7514 0.7432 0.7466 0.7480 0.7418 0.7439 0.4728 0.7493 0.748631000.0000 4.4914 cm band 6 0.9919 0.9865 0.9919 0.9864 0.9864 0.9838 0.9891 0.9811 0.9811

SSError 0.0007 0.0006 0.0005 0.0006 0.0005 0.0007 0.0355 0.0009 0.0006DegFreedom 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 4.0000 5.0000StdDev'n 0.0157 0.0140 0.0129 0.0147 0.0124 0.0148 0.1087 0.0171 0.0146R-sqAdj 0.9971 0.9977 0.9981 0.9975 0.9982 0.9974 0.8623 0.9966 0.9975R-squared 0.9983 0.9986 0.9988 0.9985 0.9989 0.9985 0.9174 0.9979 0.9985

10% Quadratic Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1326 0.1595 0.1644 0.1653 0.1603 0.1644 0.1603 0.1644 0.1622116250.0000 5.0654 cm band 2 0.2407 0.3189 0.3208 0.3198 0.3179 0.3181 0.3179 0.3208 0.318997400.0000 4.9886 cm band 3 0.2924 0.3919 0.3962 0.3902 0.3886 0.3908 0.3913 0.3935 0.391966200.0000 4.8209 cm band 4 0.3931 0.5216 0.5310 0.5257 0.5245 0.5229 0.5217 0.5202 0.527045000.0000 4.6532 cm band 5 0.5762 0.7432 0.7466 0.7480 0.7418 0.7439 0.4728 0.7493 0.748631000.0000 4.4914 cm band 6 0.8325 0.9865 0.9919 0.9864 0.9864 0.9838 0.9891 0.9811 0.981121500.0000 4.3324 cm band 714400.0000 4.1584 cm band 86500.0000 3.8129 cm band 9

SSError 0.0003 0.0004 0.0003 0.0003 0.0002 0.0003 0.0339 0.0005 0.0003DegFreedom 2.0000 2.0000 2.0000 2.0000 2.0000 2.0000 2.0000 2.0000 2.0000StdDev'n 0.0114 0.0143 0.0123 0.0116 0.0110 0.0130 0.1303 0.0164 0.0125R-sqAdj 0.9985 0.9976 0.9982 0.9984 0.9986 0.9980 0.8022 0.9969 0.9982R-squared 0.9994 0.9990 0.9993 0.9994 0.9994 0.9992 0.9209 0.9987 0.9993Tvalue 4.3027 4.3027 4.3027 4.3027 4.3027 4.3027 4.3027 4.3027 4.3027T*S 0.0489 0.0615 0.0528 0.0498 0.0472 0.0560 0.5606 0.0705 0.0536

10% Cubic Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200,000.0000 5.3010 cm band 1 0.1326 0.1595 0.1644 0.1653 0.1603 0.1644 0.1603 0.1644 0.1622 116,250.0000 5.0654 cm band 2 0.2407 0.3189 0.3208 0.3198 0.3179 0.3181 0.3179 0.3208 0.3189 97,400.0000 4.9886 cm band 3 0.2924 0.3919 0.3962 0.3902 0.3886 0.3908 0.3913 0.3935 0.3919 66,200.0000 4.8209 cm band 4 0.3931 0.5216 0.5310 0.5257 0.5245 0.5229 0.5217 0.5202 0.5270 45,000.0000 4.6532 cm band 5 0.5762 0.7432 0.7466 0.7480 0.7418 0.7439 0.4728 0.7493 0.7486 31,000.0000 4.4914 cm band 6 0.8325 0.9865 0.9919 0.9864 0.9864 0.9838 0.9891 0.9811 0.9811 21,500.0000 4.3324 cm band 714,400.0000 4.1584 cm band 86,500.0000 3.8129 cm band 9

SSError 0.0026 0.0012 0.0011 0.0014 0.0011 0.0014 0.0345 0.0016 0.0013 DegFreedom 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 StdDev'n 0.0297 0.0196 0.0189 0.0215 0.0190 0.0213 0.1073 0.0230 0.0208 R-sqAdj 0.9897 0.9955 0.9958 0.9946 0.9958 0.9947 0.8658 0.9938 0.9949 R-squared 0.9938 0.9973 0.9975 0.9968 0.9975 0.9968 0.9195 0.9963 0.9970

10% Non-Standard ModelY=ax^3+bx+c

4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3

0

1

2

3

4

LogMW

RM

RM = 19.8255 - 3.66571 LogMW

S = 0.221238 R-Sq = 96.7 % R-Sq(adj) = 95.9 %

Regression Plot

Linear

4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3

0.5

1.5

2.5

3.5

LogMW

RM

RM = 79.9215 - 28.3249 LogMW

+ 2.52197 LogMW**2

S = 0.0883707 R-Sq = 99.6 % R-Sq(adj) = 99.4 %

Regression Plot

Quadratic

4.5 4.6 4.7 4.8 4.9 5.0 5.1 5.2 5.3

0.5

1.5

2.5

3.5

LogMW

RM

RM = 579.976 - 335.455 LogMW

+ 65.2819 LogMW**2 - 4.26682 LogMW**3

S = 0.0328031 R-Sq = 100.0 % R-Sq(adj) = 99.9 %

Regression Plot

Cubic

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1301 0.1330 0.1301 0.1304 0.1304 0.1301 0.1425 0.1333 0.1330116250.0000 5.0654 cm band 2 0.2398 0.2404 0.2423 0.2379 0.2379 0.2347 0.2468 0.2436 0.243097400.0000 4.9886 cm band 3 0.2934 0.2992 0.2908 0.2890 0.2890 0.2883 0.2977 0.2897 0.294166200.0000 4.8209 cm band 4 0.3878 0.3913 0.3954 0.3913 0.3939 0.3903 0.3995 0.3949 0.393945000.0000 4.6532 cm band 5 0.5689 0.5754 0.5765 0.5754 0.5754 0.5714 0.5827 0.5795 0.580631000.0000 4.4914 cm band 6 0.8342 0.8363 0.8367 0.8286 0.8286 0.8291 0.8372 0.8308 0.831221500.0000 4.3324 cm band 7 0.9898 0.9898 0.9898 0.9898 0.9898 0.9872 0.9847 0.9897 0.9949

SSError 0.0323 0.0312 0.0301 0.0306 0.0301 0.0312 0.0306 0.0301 0.0303DegFreedom 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000 4.0000StdDev'n 0.0899 0.0884 0.0867 0.0875 0.0868 0.0883 0.0875 0.0868 0.0871R-sqAdj 0.9299 0.9323 0.9349 0.9337 0.9347 0.9324 0.9337 0.9347 0.9343R-squared 0.9533 0.9549 0.9566 0.9558 0.9565 0.9549 0.9558 0.9565 0.9562

12% Linear Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1301 0.1330 0.1301 0.1301 0.1425 0.1304 0.1304 0.1301 0.1425116250.0000 5.0654 cm band 2 0.2398 0.2404 0.2423 0.2347 0.2468 0.2379 0.2379 0.2347 0.246897400.0000 4.9886 cm band 3 0.2934 0.2992 0.2908 0.2883 0.2977 0.2890 0.2890 0.2883 0.297766200.0000 4.8209 cm band 4 0.3878 0.3913 0.3954 0.3903 0.3995 0.3913 0.3939 0.3903 0.399545000.0000 4.6532 cm band 5 0.5689 0.5754 0.5765 0.5714 0.5827 0.5754 0.5754 0.5714 0.582731000.0000 4.4914 cm band 6 0.8342 0.8363 0.8367 0.8291 0.8372 0.8286 0.8286 0.8291 0.837221500.0000 4.3324 cm band 7 0.9898 0.9898 0.9898 0.9872 0.9847 0.9898 0.9898 0.9872 0.9847

SSError 0.0076 0.0077 0.0072 0.0076 0.0081 0.0073 0.0071 0.0076 0.0081DegFreedom 3.0000 3.0000 3.0000 4.0000 5.0000 3.0000 3.0000 3.0000 3.0000StdDev'n 0.0503 0.0507 0.0488 0.0503 0.0518 0.0495 0.0487 0.0503 0.0518R-sqAdj 0.9780 0.9778 0.9793 0.9781 0.9767 0.9788 0.9795 0.9781 0.9767R-squared 0.9890 0.9889 0.9897 0.9890 0.9884 0.9894 0.9897 0.9890 0.9884

12% Quadratic Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1301 0.1330 0.1301 0.1304 0.1304 0.1301 0.1425 0.1333 0.1330116250.0000 5.0654 cm band 2 0.2398 0.2404 0.2423 0.2379 0.2379 0.2347 0.2468 0.2436 0.243097400.0000 4.9886 cm band 3 0.2934 0.2992 0.2908 0.2890 0.2890 0.2883 0.2977 0.2897 0.294166200.0000 4.8209 cm band 4 0.3878 0.3913 0.3954 0.3913 0.3939 0.3903 0.3995 0.3949 0.393945000.0000 4.6532 cm band 5 0.5689 0.5754 0.5765 0.5754 0.5754 0.5714 0.5827 0.5795 0.580631000.0000 4.4914 cm band 6 0.8342 0.8363 0.8367 0.8286 0.8286 0.8291 0.8372 0.8308 0.831221500.0000 4.3324 cm band 7 0.9898 0.9898 0.9898 0.9898 0.9898 0.9872 0.9847 0.9897 0.994914400.0000 4.1584 cm band 86500.0000 3.8129 cm band 9

SSError 0.0008 0.0010 0.0005 0.0003 0.0003 0.0004 0.0004 0.0003 0.0004DegFreedom 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000StdDev'n 0.0165 0.0181 0.0124 0.0107 0.0099 0.0116 0.0113 0.0096 0.0122R-sqAdj 0.9976 0.9972 0.9987 0.9990 0.9992 0.9988 0.9989 0.9992 0.9987R-squared 0.7508 0.7502 0.7508 0.7478 0.7480 0.7483 0.7492 0.7482 0.7478Tvalue 3.1824 3.1824 3.1824 3.1824 3.1824 3.1824 3.1824 3.1824 3.1824T*S 0.0526 0.0576 0.0394 0.0341 0.0315 0.0369 0.0361 0.0304 0.0387

12% Cubic Model

Mw LogMw GEL ID Standard-1 Standard-2 Standard-3 Standard-4 Standard-5 Standard-6 Standard-7 Standard-8 Standard-9200000.0000 5.3010 cm band 1 0.1301 0.1330 0.1301 0.1304 0.1304 0.1301 0.1425 0.1333 0.1330116250.0000 5.0654 cm band 2 0.2398 0.2404 0.2423 0.2379 0.2379 0.2347 0.2468 0.2436 0.243097400.0000 4.9886 cm band 3 0.2934 0.2992 0.2908 0.2890 0.2890 0.2883 0.2977 0.2897 0.294166200.0000 4.8209 cm band 4 0.3878 0.3913 0.3954 0.3913 0.3939 0.3903 0.3995 0.3949 0.393945000.0000 4.6532 cm band 5 0.5689 0.5754 0.5765 0.5754 0.5754 0.5714 0.5827 0.5795 0.580631000.0000 4.4914 cm band 6 0.8342 0.8363 0.8367 0.8286 0.8286 0.8291 0.8372 0.8308 0.831221500.0000 4.3324 cm band 7 0.9898 0.9898 0.9898 0.9898 0.9898 0.9872 0.9847 0.9897 0.9949

SSError 0.0024 0.0027 0.0023 0.0028 0.0027 0.0028 0.0030 0.0027 0.0027DegFreedom 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000 3.0000StdDev'n 0.0282 0.0299 0.0276 0.0306 0.0298 0.0307 0.0317 0.0303 0.0301R-sqAdj 0.9908 0.9896 0.9911 0.9891 0.9897 0.9890 0.9883 0.9893 0.9895R-squared 0.9945 0.9938 0.9947 0.9935 0.9938 0.9934 0.9930 0.9936 0.9937

12% Non-Standard Model

Conclusion

- Based on our data analysis and calculations we concluded that the cubic model fits better for 10% and 12%.

LogMW RM RM^2 RM^3 RM UkMWRM UkMW^2RMUkMW^3PFITCube PSDFCubeClimCube1ClimCub2 CICube5.301 0.1326 0.017583 0.002331 0.83722 0.7009 0.5868 4.4892 0.0118 4.438 4.54 0.102

5.0654 0.2407 0.057936 0.013945 1.33611 1.7852 2.3852 3.9963 0.3088 2.667 5.3251 2.6584.9886 0.2924 0.085498 0.025 1.50056 2.2517 3.3788 3.6375 0.5442 1.296 5.979 4.6834.8209 0.3931 0.154528 0.060745 1.88667 3.5595 6.7156 2.097 1.4849 -4.292 8.4859 12.7784.6532 0.5762 0.332006 0.191302 2.40944 5.8054 13.9878 -2.2715 3.9081 -19.086 14.5436 33.634.4914 0.8325 0.693056 0.576969 2.72167 7.4075 20.1607 -6.5836 6.1622 -33.097 19.9302 53.027

2.91944 8.5232 24.8829 -10.1215 7.9599 -44.37 24.1272 68.4983.62 13.1044 47.4379 -28.7632 17.057 -102.154 44.6272 146.781

3.725 13.8756 51.6867 -32.4985 18.8343 -113.536 48.5388 162.075

NonstFit PNonSTSDFNonSTLIM1NonSTCLIM2CINonST CubeVS.NonST CI4.4997 0.03003 4.4041 4.5952 0.1911 0.095.2423 0.26639 4.3945 6.09 1.6955 -0.9625.8612 0.41751 4.5325 7.1899 2.6574 -2.0258.2518 0.95039 5.2273 11.2764 6.0491 -6.729

14.0601 2.15919 7.1886 20.9317 13.7431 -19.88719.2372 3.20462 9.0387 29.4358 20.3971 -32.6323.2748 4.0104 10.5119 36.0377 25.5258 -42.97243.0178 7.89479 17.8931 68.1426 50.2495 -96.53146.7875 8.63044 19.3216 74.2535 54.9319 -107.143

Alpha - amylise 10%

Cubic vs Non-Satandard

LogMW RM RM^2 RM^3 RM UkMWRM UkMW^2RM UkMW^35.301 0.1326 0.017583 0.002331 0.71722 0.5144 0.3689

5.0654 0.2407 0.057936 0.013945 1.17278 1.3754 1.6134.9886 0.2924 0.085498 0.025 1.335 1.7822 2.37934.8209 0.3931 0.154528 0.060745 1.69111 2.8599 4.83634.6532 0.5762 0.332006 0.191302 1.93667 3.7507 7.26384.4914 0.8325 0.693056 0.576969 2.02778 4.1119 8.338

2.18056 4.7548 10.36822.34667 5.5068 12.9227

2.54 6.4516 16.38712.75278 7.5778 20.86

2.935 8.6142 25.28283.44333 11.8565 40.8263.57111 12.7528 45.5418

CubeFIT CubePSDFCubeCLIM1CubeCLIM2CubeCI NonSTPFIT1NonSTPSDF1NonSTCLIM1NonSTCLIM2NonST CI NonST VS. Cube4.5565 0.0114 4.5075 4.6055 0.098 4.5215 0.02056 4.456 4.5869 0.1309 -0.03284.2308 0.1518 3.5779 4.8837 1.306 4.8279 0.1546 4.3359 5.3199 0.984 0.32183.9982 0.3075 2.675 5.3214 2.646 5.2388 0.2655 4.3938 6.0837 1.6899 0.95643.0184 0.9331 -0.9964 7.0331 8.03 6.8643 0.64698 4.8054 8.9233 4.118 3.91151.8073 1.6537 -5.3081 8.9228 14.231 8.6698 1.03994 5.3602 11.9794 6.6191 7.61181.2168 1.9929 -7.3582 9.7917 17.15 9.5017 1.21644 5.6304 13.3729 7.7425 9.40730.0291 2.6591 -11.4119 11.4701 22.882 11.1103 1.5529 6.1683 16.0523 9.884 12.9979

-1.5736 3.5337 -16.7778 13.6307 30.408 13.1835 1.98011 6.8819 19.4852 12.6032 17.8052-3.8977 4.7686 -24.4155 16.6201 41.036 16.0551 2.56418 7.8947 24.2154 16.3207 24.7149-7.0961 6.425 -34.7406 20.5483 55.289 19.8317 3.32368 9.2542 30.4091 21.1549 34.134

-10.4288 8.1144 -45.3424 24.4849 69.827 23.6189 4.07881 10.6383 36.5995 25.9612 43.8661-23.069 14.3245 -84.7025 38.5645 123.267 37.1764 6.75191 15.6888 58.664 42.9752 80.2918

-27.1147 16.269 -97.1146 42.8852 140 41.3393 7.56678 17.2584 65.4202 48.1618 91.838

DTT 10%Non-standard vs Cubic

LogMW RM RM 2̂ RM 3̂ Uk RM Uk RM 2̂ UkRM 3̂5.301 0.1326 0.017583 0.002331 0.80444 0.6471 0.5206

5.0654 0.2407 0.057936 0.013945 1.01167 1.0235 1.03544.9886 0.2924 0.085498 0.025 1.275 1.6256 2.07274.8209 0.3931 0.154528 0.060745 1.49333 2.23 3.33024.6532 0.5762 0.332006 0.191302 1.605 2.576 4.13454.4914 0.8325 0.693056 0.576969 1.76278 3.1074 5.4776

1.94056 3.7658 7.30772.12667 4.5227 9.6183

2.375 5.6406 13.39652.54778 6.4912 16.53812.67278 7.1437 19.09362.79833 7.8307 21.91282.96278 8.7781 26.00742.95333 8.7222 25.75953.30444 10.9194 36.08243.45222 11.9178 41.1433.57667 12.7925 45.7547

CubePFIT CubePSDFCCubeLIM1CubeCLIM2Cube CI NonSTPFITNonSTPSDFNonSTCLIM1NonSTCLIM2NonST CI4.5074 0.01 4.4644 4.5504 0.086 4.4993 0.02551 4.4181 4.5805 0.16244.3795 0.0578 4.1308 4.6282 0.497 4.591 0.07781 4.3434 4.8387 0.49534.0962 0.2419 3.0555 5.137 2.082 5.0652 0.22033 4.364 5.7663 1.40233.6564 0.532 1.3673 5.9455 4.578 5.8294 0.41 4.5246 7.1342 2.60963.3286 0.7406 0.1423 6.515 6.373 6.3707 0.53564 4.6661 8.0754 3.40932.7176 1.1161 -2.0844 7.5197 9.604 7.3286 0.74978 4.9424 9.7147 4.77231.7838 1.6674 -5.3903 8.9579 14.348 8.7034 1.04712 5.371 12.0359 6.66480.4777 2.4096 -9.8901 10.8454 20.735 10.5114 1.42825 5.966 15.0567 9.0907-1.882 3.6996 -17.8 14.0359 31.836 13.5727 2.05971 7.0178 20.1276 13.1098

-4.0023 4.8235 -24.7563 16.7516 41.508 16.1814 2.58973 7.9398 24.4231 16.4834-5.81 5.7638 -30.6095 18.9896 49.599 18.3328 3.02317 8.7117 27.9538 19.2421

-7.8757 6.823 -37.2328 21.4814 58.714 20.7292 3.50312 9.5807 31.8777 22.297-10.9884 8.3953 -47.1105 25.1337 72.244 24.2433 4.20284 10.868 37.6186 26.7506-10.7966 8.2991 -46.5047 24.9116 71.416 24.0296 4.1604 10.7893 37.2698 26.4805-19.0866 12.3933 -72.4106 34.2374 106.648 33.0083 5.93373 14.1245 51.8921 37.7676-23.3384 14.4545 -85.5312 38.8544 124.386 37.4557 6.80664 15.7939 59.1175 43.3236-27.2991 16.3573 -97.679 43.0807 140.76 41.5276 7.60359 17.3296 65.7257 48.3961

10% Manual

Non-standard vs Cubic

NonST Vs. Cube-0.07640.00220.67921.96862.96354.83187.6834

11.644818.726125.024530.356936.417145.493644.935968.880481.062

92.3636

LogMW RM RM^2 RM^3 Uk RM Uk RM^2 Uk RM^35.301 0.1326 0.017583 0.002331 0.21778 0.0474 0.0103

5.0654 0.2407 0.057936 0.013945 0.61444 0.3775 0.2324.9886 0.2924 0.085498 0.025 0.99667 0.9933 0.994.8209 0.3931 0.154528 0.060745 1.45278 2.1106 3.06624.6532 0.5762 0.332006 0.191302 1.73944 3.0257 5.2634.4914 0.8325 0.693056 0.576969 1.91389 3.663 7.01054.3324 0.9895 0.97911 0.96883 2.02778 4.1119 8.338

2.20889 4.8792 10.77762.4 5.76 13.824

2.53278 6.415 16.24772.68833 7.2271 19.429

3.18 10.1124 32.15743.87278 14.9984 58.08553.93167 15.458 60.7757

CubePFIT1CubePSDFCubeCLIM1CubeCLIM2Cube CI NonSTPFITNonSTPSDFNonSTCLIM1NonSTCLIM2NonST CI Cube Vs. NonST5.1099 0.0069 5.088 5.132 0.0441 5.1197 0.0292 5.03873 5.2006 0.1619 0.11784.6317 0.0093 4.602 4.6614 0.0594 4.6034 0.0371 4.50026 4.7066 0.2063 0.14694.3272 0.0128 4.286 4.3681 0.0818 4.3679 0.0508 4.22681 4.5089 0.2821 0.20033.2112 0.1884 2.612 3.8107 1.199 4.6503 0.3282 3.739 5.5617 1.8227 0.62371.6073 0.4637 0.131 3.0831 2.9516 5.2569 0.6726 3.38935 7.1245 3.7352 0.78350.1413 0.7152 -2.135 2.4174 4.5521 5.8236 0.9581 3.16339 8.4839 5.3205 0.7684

-1.0538 0.9193 -3.979 1.8718 5.8512 6.2834 1.1789 3.01012 9.5566 6.5465 0.6952-3.3924 1.3164 -7.582 0.7969 8.3788 7.1725 1.5905 2.75639 11.5886 8.8322 0.4534-6.5132 1.8425 -12.377 -0.6496 11.7271 8.3378 2.1118 2.47441 14.2012 11.7268 -0.0002

-9.12 2.2792 -16.373 -1.8666 14.5068 9.2952 2.5305 2.26928 16.3211 14.0518 -0.455-12.6748 2.8716 -21.814 -3.5359 18.2777 10.5812 3.0839 2.01873 19.1437 17.125 -1.1527-27.965 5.3924 -45.126 -10.8041 34.3219 15.932 5.3251 1.14701 30.717 29.57 -4.7519

-62.1836 10.9474 -97.023 -27.3442 69.6788 27.3164 9.9535 -0.31935 54.9522 55.2716 -14.4072-65.8871 11.5442 -102.626 -29.1482 73.4778 28.518 10.4364 -0.45846 57.4946 57.953 -15.5248

Alpha - amylise 12%

Cubic vs Non-standard

CubeCLIM2 Cube CI + NonSTPFIT NonSTPSDF NonSTCLIM1 NonSTCLIM2 NonST CI Cube Vs. NonST4.6807 0.0563 4.6344 0.0367 4.53259 4.7361 0.2035 0.14724.5749 0.0694 4.4798 0.0349 4.38292 4.5767 0.1937 0.12444.4328 0.0569 4.3849 0.0369 4.2825 4.4873 0.2048 0.14794.3709 0.0796 4.3683 0.0499 4.22967 4.507 0.2773 0.19774.2806 0.1885 4.3691 0.0854 4.13192 4.6064 0.4745 0.28594.2004 0.3304 4.3939 0.1251 4.04653 4.7412 0.6947 0.36433.9737 0.8184 4.5281 0.2443 3.84968 5.2065 1.3568 0.53843.8304 1.1525 4.635 0.3183 3.75137 5.5186 1.7673 0.61483.5898 1.7273 4.8289 0.4376 3.61397 6.0439 2.4299 0.70263.3385 2.3342 5.0398 0.5566 3.4944 6.5851 3.0907 0.75653.0031 3.1448 5.3252 0.7081 3.35902 7.2913 3.9323 0.78752.7445 3.7676 5.5456 0.8205 3.26759 7.8237 4.5561 0.78852.3905 4.6165 5.8464 0.9693 3.15529 8.5376 5.3823 0.76582.0302 5.4751 6.1504 1.1158 3.05246 9.2484 6.1959 0.7208

1.43 6.8948 6.6514 1.3513 2.8996 10.4032 7.5036 0.60890.129 9.9315 7.7146 1.835 2.61976 12.8095 10.1898 0.2583

-1.2479 13.0974 8.8108 2.3195 2.37068 15.251 12.8803 -0.217-3.4203 18.0178 10.493 3.0462 2.03518 18.9509 16.9157 -1.1021-8.8361 30.0237 14.5143 4.738 1.35937 27.6693 26.3099 -3.7138

-13.4105 39.9755 17.7829 6.0866 0.8834 34.6823 33.7989 -6.1765-15.7626 45.0449 19.4308 6.7609 0.65923 38.2023 37.5431 -7.5019-18.7871 51.5257 21.5234 7.6129 0.38615 42.6606 42.2745 -9.2512-23.5512 61.6616 24.7692 8.9271 -0.01693 49.5552 49.5722 -12.0895-26.2875 67.4495 26.6097 9.6691 -0.23652 53.456 53.6925 -13.757-28.3059 71.705 27.9577 10.2113 -0.39386 56.3093 56.7032 -15.0018

DTT 12%Non-standard vs Cubic

CubePSDF CubeCLIM1CubeCLIM2Cube CI NonSTPFIT NonSTPSDFNonSTCLIM1 NonSTCLIM2NonST CI Cube Vs. NonST0.0076 4.8788 4.927 0.0483 4.944 0.0247 4.87547 5.0126 0.1371 -0.08880.0078 4.7136 4.7635 0.0499 4.7561 0.0322 4.66662 4.8456 0.179 -0.12920.0094 4.6008 4.6604 0.0596 4.6017 0.0372 4.49851 4.7049 0.2064 -0.14680.0104 4.5523 4.6186 0.0663 4.5366 0.0369 4.43426 4.639 0.2047 -0.13840.0104 4.4604 4.5265 0.0662 4.4346 0.0327 4.34381 4.5254 0.1816 -0.11540.0154 4.2522 4.3499 0.0977 4.3656 0.0567 4.20816 4.5231 0.3149 -0.21720.0817 3.5874 4.1076 0.5203 4.4408 0.1737 3.95837 4.9232 0.9648 -0.44450.1057 3.3652 4.038 0.6728 4.4841 0.2106 3.89946 5.0687 1.1692 -0.49640.1762 2.7223 3.8436 1.1213 4.6248 0.3115 3.75982 5.4897 1.7299 -0.60860.3288 1.3453 3.4383 2.0929 4.9555 0.51 3.53959 6.3714 2.8318 -0.73880.4596 0.1687 3.094 2.9253 5.2476 0.6678 3.39357 7.1017 3.7081 -0.78280.8056 -2.9515 2.1762 5.1278 6.0275 1.0569 3.09295 8.9621 5.8691 -0.74140.9798 -4.5276 1.709 6.2366 6.4194 1.243 2.96819 9.8707 6.9025 -0.66591.1961 -6.4888 1.1243 7.6131 6.904 1.4679 2.8285 10.9795 8.151 -0.53791.4721 -8.9984 0.3713 9.3697 7.5188 1.7472 2.66787 12.3698 9.7019 -0.33221.7969 -11.9607 -0.5235 11.4372 8.2375 2.0675 2.4971 13.9778 11.4807 -0.04362.4516 -17.9545 -2.3503 15.6042 9.6709 2.6931 2.19353 17.1482 14.9547 0.64952.8945 -22.0239 -3.6007 18.4232 10.6306 3.105 2.00957 19.2516 17.242 1.18123.2617 -25.4053 -4.6449 20.7604 11.421 3.4412 1.86661 20.9753 19.1087 1.65173.9945 -32.1702 -6.7457 25.4246 12.9857 4.1003 1.6012 24.3701 22.7689 2.65575.001 -41.4933 -9.6621 31.8312 15.1117 4.9858 1.26865 28.9547 27.6861 4.1451

5.9832 -50.6179 -12.5356 38.0823 17.1647 5.8328 0.96992 33.3595 32.3896 5.69277.3837 -63.6677 -16.6713 46.9964 20.0625 7.0186 0.57554 39.5494 38.9739 8.02268.4489 -73.618 -19.8416 53.7764 22.2468 7.9066 0.29434 44.1992 43.9049 9.87159.576 -84.1658 -23.2156 60.9501 24.5423 8.8355 0.01055 49.074 49.0635 11.8867

11.0472 -97.9603 -27.6458 70.3145 27.5177 10.0345 -0.3428 55.3783 55.7211 14.5934

12% Manual

Cubic vs Non-standard

Conclusion

- In addition the DTT process of removal of the Vitelline envelope is better than the α-Amylise process for both the Non-standard and cubic models in both 10% and 12% gels.

Analysis of the confidence intervals produced by the cubic and non-standard models showed the

non-standard model to work better for the 10% gel and the cubic to work better for the 12% gel.