[BioC] error message----Estimation of var.prior failed

Jia Liu jliu at tigr.ORG
Fri Jun 1 21:42:05 CEST 2007


Hello All,

I am trying to fit the data to the linear model to get the differential 
expressed gene list.  The experiment has a loop design with 12 
hybridizations consisting of 6 samples (each with 4 biological 
replicates). After I run the following code, i encountered the warning 
message from "eBayes" function. Then when i run the topTable function, 
the resulting table contains duplicated entries as shown below. I also 
enclosed the "targets", "design" and the MA object "MA.AB" for your 
information. Can someone help me idntify the problem? Thanks a lot!

Jia


 >design.names=list(c("slide1","slide2","slide3","slide4","slide5","slide6","slide7","slide8","slide9","slide10","slide11","slide12"),c("DB-DM","DM-NM","DM-DT","NM-NB","NT-NM"))
 >design=matrix(c(1,1,0,1,0,1,0,0,0,0,0,1,0,0,0,0,0,1,0,0,0,1,-1,0,-1,-1,0,-1,0,0,-1,-1,0,-1,0,0,0,0,-1,-1,0,-1,0,0,0,0,0,0,1,0,0,-1,1,0,1,0,0,0,0,1),12,5,byrow=T,dimnames=design.names)
 > fit<-lmFit(MA.AB,design=design,weights=MA.AB$weights, 
p.adjust="Hochberg")
 > fit<-eBayes(fit)
*Warning message:
Estimation of var.prior failed - set to default value in: ebayes(fit = 
fit, proportion = proportion, stdev.coef.lim = stdev.coef.lim)*

 > topTable(fit[,1])      

Block Row Column         ID     Name Status
4080      7  21      7 MZ00043373 TC220219   gene
32550     7  21      7 MZ00043373 TC220219   gene
13125    22  24      5 MZ00028432 TC225126   gene
41595    22  24      5 MZ00028432 TC225126   gene
23889    40  22     16 MZ00027664 TC225103   gene
52359    40  22     16 MZ00027664 TC225103   gene
15428    26  20      1 MZ00027244 TC239087   gene
43898    26  20      1 MZ00027244 TC239087   gene
27756    47  21      8 MZ00043374 TC206176   gene
56226    47  21      8 MZ00043374 TC206176   gene
                                     Annotation
4080                                                                                                      
cellulose synthase {Oryza sativa (japonica cultivar-group);}
32550                                                                                                     
cellulose synthase {Oryza sativa (japonica cultivar-group);}
13125 hypothetical protein {Oryza sativa (japonica cultivar-group);} 
^|^GB|BAB92324.1|20804634|AP003253 hypothetical protein {Oryza sativa 
(japonica cultivar-group);}
41595 hypothetical protein {Oryza sativa (japonica cultivar-group);} 
^|^GB|BAB92324.1|20804634|AP003253 hypothetical protein {Oryza sativa 
(japonica cultivar-group);}
23889               putative CER1 {Oryza sativa (japonica 
cultivar-group);} ^|^GB|AAG21908.1|10716610|AC026815 putative CER1 
{Oryza sativa (japonica cultivar-group);}
52359               putative CER1 {Oryza sativa (japonica 
cultivar-group);} ^|^GB|AAG21908.1|10716610|AC026815 putative CER1 
{Oryza sativa (japonica cultivar-group);}
15428        OSJNBa0029H02.22 {Oryza sativa (japonica cultivar-group);} 
^|^GB|CAD41488.1|21740509|OSJN00025 OSJNBa0029H02.27 {Oryza sativa 
(japonica cultivar-group);}
43898        OSJNBa0029H02.22 {Oryza sativa (japonica cultivar-group);} 
^|^GB|CAD41488.1|21740509|OSJN00025 OSJNBa0029H02.27 {Oryza sativa 
(japonica cultivar-group);}
27756               cellulose synthase-6 {Zea mays;}
56226               cellulose synthase-6 {Zea mays;}
          logFC   AveExpr         t      P.Value adj.P.Val            B
4080  -2.446885  9.317316 -8.446857 2.037268e-05 0.2538640  2.079626317
32550 -2.446885  9.317316 -8.446857 2.037268e-05 0.2538640  2.079626317
13125  1.913508 11.108700  5.139045 2.808850e-04 0.5331159  0.539694800
41595  1.913508 11.108700  5.139045 2.808850e-04 0.5331159  0.539694800
23889 -1.624284 11.770228 -5.012985 3.443437e-04 0.5331159  0.372559874
52359 -1.624284 11.770228 -5.012985 3.443437e-04 0.5331159  0.372559874
15428 -1.456442  9.134642 -5.143359 5.125077e-04 0.5331159 -0.006356757
43898 -1.456442  9.134642 -5.143359 5.125077e-04 0.5331159 -0.006356757
27756 -2.108918  8.578181 -6.546458 4.367518e-04 0.5331159 -0.091344144
56226 -2.108918  8.578181 -6.546458 4.367518e-04 0.5331159 -0.091344144

 > targets
   SlideNumber FileName Cy3 Cy5
1            1 A160.gpr  DB  NB
2            2 A164.gpr  DB  DM
3            3 A165.gpr  DM  NM
4            4 A166.gpr  DM  DT
5            5 A167.gpr  DT  NT
6            6 A168.gpr  DT  DB
7            7 A169.gpr  NB  DB
8            8 A170.gpr  NB  NT
9            9 A171.gpr  NM  DM
10          10 A077.gpr  NM  NB
11          11 A078.gpr  NT  DT
12          12 A079.gpr  NT  NM


 > design
        DB-DM DM-NM DM-DT NM-NB NT-NM
slide1      1     1     0     1     0
slide2      1     0     0     0     0
slide3      0     1     0     0     0
slide4      0     0     1     0     0
slide5      0     1    -1     0    -1
slide6     -1     0    -1     0     0
slide7     -1    -1     0    -1     0
slide8      0     0     0    -1    -1
slide9      0    -1     0     0     0
slide10     0     0     0     1     0
slide11     0    -1     1     0     1
slide12     0     0     0     0     1
 > MA.AB
An object of class "MAList"
$weights
     B167 B168 B169 B170 B177 B180 B181 B182 B183 B070 B072 B073
[1,]    0    0    0    0    0    0    0    0    0    0    0    0
[2,]    0    0    0    0    0    0    0    0    0    0    0    0
[3,]    0    0    0    0    0    0    0    0    0    0    0    0
[4,]    0    0    0    0    0    0    0    0    0    0    0    0
[5,]    0    1    1    1    1    1    0    1    0    0    1    1
56935 more rows ...

$targets
  SlideNumber FileName Cy3 Cy5
1           1 B167.gpr  DB  NB
2           2 B168.gpr  DB  DM
3           3 B169.gpr  DM  NM
4           4 B170.gpr  DM  DT
5           5 B177.gpr  DT  NT
7 more rows ...

$genes
  Block Row Column         ID        Name Status
1     1   1      1 MZ00002380  AZM4_14521   gene
2     1   1      2 MZ00002386  AZM4_49988   gene
3     1   1      3 MZ00002404 AZM4_100937   gene
4     1   1      4 MZ00002410 AZM4_135577   gene
5     1   1      5 MZ00002428  AZM4_79828   gene
                                                                                                        
Annotation
1                                   phosphatidylinositol 3,5-kinase-like 
{Oryza sativa (japonica cultivar-group);}
2                                                        OSJNBa0087O24.3 
{Oryza sativa (japonica cultivar-group);}
3 hypothetical protein similar to Oryza sativa chromosome 1, P0431G06.13 
{Oryza sativa (japonica cultivar-group);}
4                                                   putative phosphatase 
{Oryza sativa (japonica cultivar-group);}
5                              putative flavonol 3-O-glucosyltransferase 
{Oryza sativa (japonica cultivar-group);}
56935 more rows ...

$source
[1] "genepix"

$printer
$ngrid.r
[1] 12

$ngrid.c
[1] 4

$nspot.r
[1] 26

$nspot.c
[1] 26

attr(,"class")
[1] "PrintLayout"

$M
           B167       B168        B169       B170       B177       B180
[1,] -1.3779913 -1.8833111 -1.62708741 -1.3495565 -1.1180732 -0.5628514
[2,] -0.4571145 -0.6198667 -0.06527221 -0.5730541 -0.6867249 -0.6093460
[3,] -0.2940607 -0.2293377 -0.20736120 -0.9225977 -1.0349128 -0.8617654
[4,] -1.4274691 -1.1814338 -1.45430688 -0.5903722 -0.8967976 -1.2961053
[5,]  0.9068494  0.1585617 -0.10686769  0.3406360 -0.5760433 -1.0153630
           B181       B182       B183      B070        B072       B073
[1,]         NA -1.8638750 -0.9362995 -2.577714 -1.99677057 -0.1696838
[2,] -1.2031207 -0.1840673 -0.9566250 -2.154500 -0.52701635 -0.2899944
[3,]  0.2947026  1.2401698         NA -1.686424 -0.01693860 -0.1007202
[4,] -1.4675358 -2.6755268         NA -2.531036 -0.69658202 -2.5460154
[5,]  1.0599882  1.4856234  1.4375392 -1.177492  0.49266749  1.4147140
56935 more rows ...

$A
         B167     B168     B169     B170     B177     B180     B181     B182
[1,] 7.633485 6.241182 6.675137 6.402408 6.370169 6.356133       NA 5.784979
[2,] 8.937003 6.628853 5.813245 6.034256 6.138358 5.515603 5.656917 5.668056
[3,] 5.512734 6.183253 6.367349 5.099702 5.612979 5.124473 5.581835 4.329897
[4,] 7.185585 6.401859 5.634371 6.137675 6.210610 6.660527 5.355347 5.621532
[5,] 7.074069 8.971200 9.019419 9.814719 9.862116 8.833574 7.790254 8.648343
         B183     B070     B072     B073
[1,] 6.905622 6.242238 5.785340 6.237580
[2,] 7.115129 6.069744 5.617608 6.084069
[3,]       NA 5.322445 5.309317 5.177219
[4,]       NA 6.566366 5.293450 4.902802
[5,] 8.803978 5.990159 8.700017 7.253411
56935 more rows ...



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