I am trying to do ttest for each plate which has equal number of disease and
controls. by searching this forum I found one posting suggesting OP to use
"by(eo,PlateID, function(.sub) t.test(IL1Ra~Group,data=.sub))". when i
modified this for my use I used to get the pvalues for each plate, recently
upgraded to R2.9.x and now i am getting following error when i use this 
> by(eo,PlateID, function(.sub) t.test(IL1Ra~Group,data=.sub))
Error in t.test.formula(IL1Ra ~ Group, data = .sub) : 
  grouping factor must have exactly 2 levels

I checked my data file and the plate ID has "P16-P47" and the disease column
is "Y/N". I have search all but could not locate the answer. Can anyone
suggest me the direction where i am going wrong. 
for more detail i amm posting a part of my data.

SampleID        PlateID Sex     Disease Duration        Age     prt1    prt2    
prt3    prt4
1       P16     F       N               25.33282811     836.08979       
20.04582692     295.74  11731.43
2       P16     F       N               32.74912883     243.0652116     
53.16487056     383.63  4451.36
3       P16     M       N               3.49961999      181.5587298     
23.24604762     522.52  325.52
4       P16     M       N               5.249771666     261.0978097     
19.69024684     833.61  2229.04
5       P16     M       N               39.16612385     237.334794      
83.97284692     694.03  1204.36
6       P16     F       N               33.49929145     540.486536      
29.69569346     895.36  72105.75
7       P16     F       N               3.915997361     5215.378446     
35.44324704     1023.63 101680.69
8       P16     F       N               33.49929145     466.5188732     
12.93422535     814.26  7169.61
9       P16     F       N               38.1661348      706.5117791     
19.69024684     331.62  7296.72
10      P16     M       N               0.916030215     242.3011559     
36.40117263     90.1    35962.08
11      P16     F       Y       14975.25        42.08281552     133.8052501     
29.0570764      260.55  1280.29
12      P16     M       Y       1095.75 40.41593932     247.6495456     
19.69024684     161.64  6685.6
13      P16     F       Y       2922    25.08300169     231.6043765     
9.022844487     902.53  37571.39
14      P16     F       Y       12053.25        37.49924765     158.6370595     
24.90606549     1.63    471.15
15      P16     F       Y       14610   76.58209602     278.3837309     
41.51010914     152.13  16285.49
16      P16     F       Y       3287.25 49.58239171     128.4568603     
17.91234645     274.99  41823.21
17      P17     M       N               2.333079994     397.4440508     
55.20987654     366.37  90011.27
18      P17     F       N               4.749435461     222.0839813     
73.62489675     271.65  903.89
19      P17     F       N               1.749468315     676.8904636     
47.7037037      721.19  663.15
20      P17     M       N               33.24946503     413.712486      
27.42386831     611.64  4195.86
21      P17     F       N               15.4162131      346.9913086     
30.71604938     302.29  2151.47
22      P17     F       N               33.3327405      2598.071161     
107.8088094     1800.44 102005.12
23      P17     M       N               9.832656179     3296.535581     
54.1563786      2642.73 676.78
24      P17     F       N               3.749446413     488.2589867     
15.29151952     562.88  11619.62
25      P17     M       N               9.749380705     159.1536586     
23.41690259     132.64  931.14
26      P17     M       Y       12053.25        50.91548265     173.7764722     
18.90280088     52.25   15962.05
27      P17     M       Y       3652.5  25.83248096     264.8254141     
35.32510288     497.33  15504.95
28      P17     F       Y               28.41607073     415.3599225     
35.0617284      1600.67 14224.39
29      P17     F       Y       6209.25 60.74882219     213.7280879     
44.41152263     111.36  9018.48
30      P17     M       Y       4748.25 47.33258719     264.0016959     
31.24279835     366.81  3809.83
31      P17     M       Y       16071   54.49906148     112.4128794     
34.40329218     114.56  1727.02
32      P17     M       Y       7670.25 52.33253243     145.5753317     
32.2962963      44.92   15264.84
33      P17     M       Y       8766    31.74913978     220.2561296     
34.40329218     130.31  2377.36
34      P17     F       Y       8400.75 40.83231669     194.6662059     
48.09876543     407.83  19587.61
35      P17     M       Y       12053.25        36.1661567      212.1613578     
96.51775947     621.42  3269.9
36      P17     M       Y       8035.5  57.91540599     115.2852178     
24.79012346     282.88  8960.27
37      P17     M       Y       5478.75 36.33270765     160.4592669     
40.72427984     102.32  5006.77
38      P17     F       Y       9496.5  34.74910692     329.8991551     
40.32921811     417.32  248857.58
39      P18     F       N               14.2496731      538.5430144     
54.34303351     396     4432.94
40      P18     M       N               11.83263428     236.8218238     
48.37595532     148.94  1414.11
41      P18     M       N               7.582851659     322.2342976     
87.83185053     280.84  16294.98
42      P18     F       N               8.83266713      288.4507237     
30.47472075     256.38  348.03
43      P18     M       N               43.58245741     297.1312253     
3.276042927     368.22  3627.89
44      P18     F       N               45.49916004     297.1312253     
160.4104035     635.12  1325.33
45      P18     M       N               10.83264523     179.2726374     
39.85155791     64.24   763.59
46      P18     M       N               18.5827312      3389.23762      
71.66394523     1105.37 45291.49
47      P18     F       N               7.416300711     1345.377837     
29.62228101     1195.82 19241.32
48      P18     M       Y       4383    19.16634287     137.1490061     
20.22826506     157.45  4026.95
49      P18     M       Y       4748.25 19.0830674      226.7358839     
93.61453199     507.45  1274.6
50      P18     M       Y       4748.25 19.0830674      285.870034      
60.87840486     330.91  586.98
51      P18     M       Y       3652.5  12.33297048     195.6870101     
39.56741133     48.1    16561.4
52      P18     F       Y       4017.75 11.24970596     145.4550742     
42.55095042     20.56   732.42
53      P18     F       Y       1461    3.999956195     385.813107      
63.1515775      204.62  2347.46
54      P18     M       Y       4748.25 16.08310025     117.965944      
58.03693906     78.33   2709.32
55      P18     M       Y       1826.25 13.99984668     175.9106574     
67.41377621     65.53   2069.11
56      P18     M       Y       730.5   14.9158769      89.68575957     
27.91740153     22.62   389.59
57      P18     F       Y       6209.25 17.4161912      191.9295031     
32.60582011     127.63  20291.34
58      P18     F       Y       365.25  5.249771666     121.723451      
61.7308446      49.39   11463.27
59      P18     F       Y       4748.25 18.33290477     179.0748739     
53.20644719     46.16   111361.43
60      P18     M       Y       1826.25 6.083209766     330.6801911     
27.06496179     489.04  12258.96
61      P18     F       Y       3287.25 17.58274215     182.8323809     
29.48020772     115.1   15324.67
62      P18     F       Y       3652.5  44.16606909     248.4898719     
87.24178099     139.33  16443.86
63      P19     M       N               37.99958385     2197.263844     
86.43305867     1492.06 3547.88
64      P19     F       N               37.58252312     4588.403833     
152.9115464     1722.46 9243.22
65      P19     F       N               34.91565787     316.0623502     
17.19804354     1582.79 12217.62
66      P19     F       N               42.74901931     213.1054131     
32.99689553     212.94  2714.66
67      P19     F       N               10.08316596     4715.658374     
50.91906722     1543.61 2490.68
68      P19     F       N               43.33263099     2.258320699     
0.524578302     561.12  5985.07
69      P19     F       Y       365.25  8.999901438     208.5132426     
32.8293986      45.72   2051.68
70      P19     M       Y       4748.25 12.83262333     0       0       192.51  
1255.58
71      P19     M       Y       1461    16.83257952     218.2716049     
65.82629413     255.01  4486.39
72      P19     M       Y       5844    17.4161912      102.60631       
27.80449065     49.77   622.01
73      P19     M       Y       4748.25 18.33290477     182.9692941     
9.965221488     159.66  2392.13
74      P19     F       Y       730.5   17.24964025     100.8842461     
27.97198758     157.25  176209.9
75      P19     M       Y       4383    16.41620215     0.098187856     0       
171.67  65286.23
76      P19     M       Y       1461    6.16648524      962.1232959     
112.1204345     81.67   870.82



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