[BioC] Incorporating direct arrays to reference design

mjonczyk at biol.uw.edu.pl mjonczyk at biol.uw.edu.pl
Thu Aug 25 08:25:45 CEST 2011


Dear Gordon & List Members,

I've made few hybridizations which I'd like to incorporate to the design
described in thread "Strange results in contrasts with dye-swap" (It was
reference design, two temperature treatments
with seven samples (time points) in both, all compared directly to "time 0" sample).

Treatments I'd like to merge with that experiment are direct comparison of:

c2 vs k2
c6 vs k6
c8 vs k8

As main experiment this comparisons were repeated four times with dye-swap (two
labellings in both directions), using the same biological material as in
corresponding replications in the main experiment.

Here is my new target frame:

FileName	Cy3	Cy5	powt
_024_1_vs_7ex.gpr	k0	k2	1
_228_k0_vs_16ex.gpr	k0	c12	1
_229_k0_vs_8ex.gpr	k0	c4	1
_230_k0_vs_15ex.gpr	k0	k10	1
_235_k0_vs_19ex.gpr	k0	k14	1
_237_k0_vs_14ex.gpr	k0	c10	1
_238_k0_vs_10ex.gpr	k0	c6	1
_239_k0_vs_12ex.gpr	k0	c8	1
_240_k0_vs_13ex.gpr	k0	k8	1
_248_k0_vs_17ex.gpr	k0	k12	1
_249_k0_vs_9ex.gpr	k0	k4	1
_253_k0_vs_11ex.gpr	k0	k6	1
_254_k0_vs_6ex.gpr	k0	c2	1
_256_k0_vs_18ex.gpr	k0	c14	1
_189_K8_vs_K0ex.gpr	k8	k0	2
_190_K4_vs_K0ex.gpr	k4	k0	2
_191_K6_vs_K0ex.gpr	k6	k0	2
_192_C8_vs_K0ex.gpr	c8	k0	2
_193_C10_vs_K0ex.gpr	c10	k0	2
_194_C2_vs_K0ex.gpr	c2	k0	2
_198_C14_vs_K0ex.gpr	c14	k0	2
_200_K12_vs_K0ex.gpr	k12	k0	2
_202_K10_vs_K0ex.gpr	k10	k0	2
_206_C6_vs_K0ex.gpr	c6	k0	2
_207_C12_vs_K0ex.gpr	c12	k0	2
_208_K2_vs_K0ex.gpr	k2	k0	2
_209_K14_vs_K0ex.gpr	k14	k0	2
_210_C4_vs_K0ex.gpr	c4	k0	2
_201_k0_vs_C2ex.gpr	k0	c2	3
_203_k0_vs_K6ex.gpr	k0	k6	3
_205_k0_vs_C12ex.gpr	k0	c12	3
_217_k0_vs_K14ex.gpr	k0	k14	3
_219_k0_vs_C4ex.gpr	k0	c4	3
_231_k0_vs_C10ex.gpr	k0	c10	3
_232_k0_vs_K8ex.gpr	k0	k8	3
_233_k0_vs_C8ex.gpr	k0	c8	3
_234_k0_vs_C14ex.gpr	k0	c14	3
_235_k0_vs_C6ex.gpr	k0	c6	3
_236_k0_vs_K10ex.gpr	k0	k10	3
_237_k0_vs_K2ex.gpr	k0	k2	3
_238_k0_vs_K4ex.gpr	k0	k4	3
_239_k0_vs_K12ex.gpr	k0	k12	3
_004_8_vs_1ex.gpr	c8	k0	4
_005_10_vs_1ex.gpr	c10	k0	4
_006_2_vs_1ex.gpr	c2	k0	4
_007_15_vs_1ex.gpr	k14	k0	4
_008_4_vs_1ex.gpr	c4	k0	4
_010_3_vs_1ex.gpr	k2	k0	4
_011_9_vs_1ex.gpr	k8	k0	4
_012_14_vs_1ex.gpr	c14	k0	4
_014_12_vs_1ex.gpr	c12	k0	4
_026_13_vs_1ex.gpr	k12	k0	4
_036_11_vs_1ex.gpr	k10	k0	4
_037_6_vs_1ex.gpr	c6	k0	4
_043_7_vs_1ex.gpr	k6	k0	4
_044_5_vs_1ex.gpr	k4	k0	4
127	k2	c2	1
049	c2	k2	2
133	c2	k2	4
128	k2	c2	3
047	k6	c6	1
050	c6	k6	2
046	c6	k6	4
048	k6	c6	3
130	k8	c8	1
134	c8	k8	2
132	c8	k8	4
045	k8	c8	3

So design matrix will be
> xdesign=modelMatrix(xtrg,ref="k0")
> xdesign
      c10 c12 c14 c2 c4 c6 c8 k10 k12 k14 k2 k4 k6 k8
 [1,]   0   0   0  0  0  0  0   0   0   0  1  0  0  0
 [2,]   0   1   0  0  0  0  0   0   0   0  0  0  0  0
 [3,]   0   0   0  0  1  0  0   0   0   0  0  0  0  0
 [4,]   0   0   0  0  0  0  0   1   0   0  0  0  0  0
 [5,]   0   0   0  0  0  0  0   0   0   1  0  0  0  0
 [6,]   1   0   0  0  0  0  0   0   0   0  0  0  0  0
 [7,]   0   0   0  0  0  1  0   0   0   0  0  0  0  0
 [8,]   0   0   0  0  0  0  1   0   0   0  0  0  0  0
 [9,]   0   0   0  0  0  0  0   0   0   0  0  0  0  1
[10,]   0   0   0  0  0  0  0   0   1   0  0  0  0  0
[11,]   0   0   0  0  0  0  0   0   0   0  0  1  0  0
[12,]   0   0   0  0  0  0  0   0   0   0  0  0  1  0
[13,]   0   0   0  1  0  0  0   0   0   0  0  0  0  0
[14,]   0   0   1  0  0  0  0   0   0   0  0  0  0  0
[15,]   0   0   0  0  0  0  0   0   0   0  0  0  0 -1
[16,]   0   0   0  0  0  0  0   0   0   0  0 -1  0  0
[17,]   0   0   0  0  0  0  0   0   0   0  0  0 -1  0
[18,]   0   0   0  0  0  0 -1   0   0   0  0  0  0  0
[19,]  -1   0   0  0  0  0  0   0   0   0  0  0  0  0
[20,]   0   0   0 -1  0  0  0   0   0   0  0  0  0  0
[21,]   0   0  -1  0  0  0  0   0   0   0  0  0  0  0
[22,]   0   0   0  0  0  0  0   0  -1   0  0  0  0  0
[23,]   0   0   0  0  0  0  0  -1   0   0  0  0  0  0
[24,]   0   0   0  0  0 -1  0   0   0   0  0  0  0  0
[25,]   0  -1   0  0  0  0  0   0   0   0  0  0  0  0
[26,]   0   0   0  0  0  0  0   0   0   0 -1  0  0  0
[27,]   0   0   0  0  0  0  0   0   0  -1  0  0  0  0
[28,]   0   0   0  0 -1  0  0   0   0   0  0  0  0  0
[29,]   0   0   0  1  0  0  0   0   0   0  0  0  0  0
[30,]   0   0   0  0  0  0  0   0   0   0  0  0  1  0
[31,]   0   1   0  0  0  0  0   0   0   0  0  0  0  0
[32,]   0   0   0  0  0  0  0   0   0   1  0  0  0  0
[33,]   0   0   0  0  1  0  0   0   0   0  0  0  0  0
[34,]   1   0   0  0  0  0  0   0   0   0  0  0  0  0
[35,]   0   0   0  0  0  0  0   0   0   0  0  0  0  1
[36,]   0   0   0  0  0  0  1   0   0   0  0  0  0  0
[37,]   0   0   1  0  0  0  0   0   0   0  0  0  0  0
[38,]   0   0   0  0  0  1  0   0   0   0  0  0  0  0
[39,]   0   0   0  0  0  0  0   1   0   0  0  0  0  0
[40,]   0   0   0  0  0  0  0   0   0   0  1  0  0  0
[41,]   0   0   0  0  0  0  0   0   0   0  0  1  0  0
[42,]   0   0   0  0  0  0  0   0   1   0  0  0  0  0
[43,]   0   0   0  0  0  0 -1   0   0   0  0  0  0  0
[44,]  -1   0   0  0  0  0  0   0   0   0  0  0  0  0
[45,]   0   0   0 -1  0  0  0   0   0   0  0  0  0  0
[46,]   0   0   0  0  0  0  0   0   0  -1  0  0  0  0
[47,]   0   0   0  0 -1  0  0   0   0   0  0  0  0  0
[48,]   0   0   0  0  0  0  0   0   0   0 -1  0  0  0
[49,]   0   0   0  0  0  0  0   0   0   0  0  0  0 -1
[50,]   0   0  -1  0  0  0  0   0   0   0  0  0  0  0
[51,]   0  -1   0  0  0  0  0   0   0   0  0  0  0  0
[52,]   0   0   0  0  0  0  0   0  -1   0  0  0  0  0
[53,]   0   0   0  0  0  0  0  -1   0   0  0  0  0  0
[54,]   0   0   0  0  0 -1  0   0   0   0  0  0  0  0
[55,]   0   0   0  0  0  0  0   0   0   0  0  0 -1  0
[56,]   0   0   0  0  0  0  0   0   0   0  0 -1  0  0
[57,]   0   0   0  1  0  0  0   0   0   0 -1  0  0  0
[58,]   0   0   0 -1  0  0  0   0   0   0  1  0  0  0
[59,]   0   0   0 -1  0  0  0   0   0   0  1  0  0  0
[60,]   0   0   0  1  0  0  0   0   0   0 -1  0  0  0
[61,]   0   0   0  0  0  1  0   0   0   0  0  0 -1  0
[62,]   0   0   0  0  0 -1  0   0   0   0  0  0  1  0
[63,]   0   0   0  0  0 -1  0   0   0   0  0  0  1  0
[64,]   0   0   0  0  0  1  0   0   0   0  0  0 -1  0
[65,]   0   0   0  0  0  0  1   0   0   0  0  0  0 -1
[66,]   0   0   0  0  0  0 -1   0   0   0  0  0  0  1
[67,]   0   0   0  0  0  0 -1   0   0   0  0  0  0  1
[68,]   0   0   0  0  0  0  1   0   0   0  0  0  0 -1

*Is* it correct? I.e. is it make correct use of direct (cx vs kx) comparisons?

*Can* I use the same code as previously

> xdesign_dye=cbind(DyeEffect=1,xdesign)
> fit.userguide.dye=lmFit(img_lA_ncav,xdesign_dye)
>
contrast.matrix.d=makeContrasts(p2="c2-k2",p4="c4-k4",p6="c6-k6",p8="c8-k8",p10="c10-k10",p12="c12-k12",p14="c14-k14",levels=xdesign_dye)
> fit2.ug.d=contrasts.fit(fit.userguide.dye,contrast.matrix.d)
> fit2.ug.d=eBayes(fit2.ug.d)
> test.ug.d=decideTests(fit2.ug.d,method="global",adjust.method="BH",p.value=0.05)

Thanks in advance,
Best Regards,

Maciej Jończyk, MSc
Department of Plant Molecular Ecophysiology
Institute of Plant Experimental Biology
Faculty of Biology, University of Warsaw
02-096 Warszawa, Miecznikowa
1



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