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$Id$ |
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== Introduction == |
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There are many plug-ins in the Illumina plug-in package for BASE. This |
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file gives detailed information on some of the contributed plug-ins. |
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== Illumina expression background correction plug-in == |
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This plug-in will remove a per-slide global background from all |
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spots. The background is calculated from a set of negative control |
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spot on the array. See implementation details below for details. |
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|
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=== Parameters === |
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There is one parameter to set that specifies how background |
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intensities should be calculated. Allowed values are median or mean, |
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i.e. the background is either the median or the mean of the negative |
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control spots on the array. |
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The expression of the background probes is optionally saved to a |
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file. The default is not to save the expression matrix but this can be |
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changed during job configuration. |
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=== Implementation details === |
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Each assay is treated separately, i.e., no samples are combined |
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together. All calculations are made on the current bioassay data |
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implying that this plug-in should be used early in analysis and before |
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background spots are removed. |
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A spot is considered to be a negative control spot if it has a |
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''Control group name'' exactly matching the string ''negative''. |
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|
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== Illumina detection P-value calculation == |
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This plug-in implements !BeadStudio like detection P-value calculations |
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for Illumina expression data (see |
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http://www.genomecenter.ucdavis.edu/expression_analysis/documents/illumina_normalization_081201.pdf) |
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on detection P-values. |
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The plug-in will ''always'' base the detection P-value calculation on |
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raw data values, ''i.e.'', the mean raw intensity for the different |
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signals. By default the calculations are based on negative controls |
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available in the root bioassay set for the current analysis |
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branch. The user may change this to only use negative controls in the |
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current bioassay set. |
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The detection P-value plug-in does not filter the assays, it provides |
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the detection P-values usable in a filter step after running this |
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plug-in. |
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=== Parameters === |
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The plug-in requires input of array type since detection P-value are |
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calculated differently depending on array type. |
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Users may select to use negative controls in the current bioassay set |
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only. The default behaviour is to use all negative controls in the |
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root bioassay set for the current bioassay set. |
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A cut off parameter is available to exclude outliers within |
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the negative controls. The `cutoff` defines the acceptable negative |
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control signal range |
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{{{ |
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median-MAD*cutoff < I < median+MAD*cutoff |
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}}} |
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where `MAD` is the median absolute deviation. |
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=== Implementation details === |
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|
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Each assay is treated separately, i.e., no samples are combined |
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together. All calculations are made on the raw bead-type level data, |
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i.e., on the average expression value for each bead type and raw data |
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is always used irrespective when in analysis the detection P-value is |
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calculated. |
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''Pvalue calculation for whole genome arrays:'' |
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For all signals `i` calculate the detection P-value as |
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`Pvalue = 1-R/N` where `R` is the rank of the signal `i` relative to the |
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negative controls and `N` is the number of negative controls. |
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''Pvalue calculation for others array types (DASL, miRNA, !VeraCode |
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DASL, and Focused Arrays):'' |
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For all signals `i` calculate the detection P-value as |
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`Pvalue = 1/2 - 1/2 * erf( [i-AvgControl]/StdControl/sqrt(2) )` where |
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`AvgControl` is the average intensity of the negative controls, |
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`StdControl` is the standard deviation of the the negative controls, |
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and `erf` is the error function |
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(http://mathworld.wolfram.com/Erf.html). The error function is used |
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for arguments within the range (-4,4). To save CPU cycles, the function |
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value for arguments outside this range is set to -1 and 1, |
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respectively. |
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|
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A spot is considered to be a negative control spot if it has an |
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''Control group name'' exactly matching the string ''negative''. |
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== Control Summary plots == |
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This extension provides overview plots for Illumina expression data. The |
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'Overview plots' tab becomes available in the experiment analysis tree |
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when the user selects a bioassay set. The plots are automatically |
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generated when the user clicks on the Overview plots tab. The display |
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of the plots cannot be changed by the user and the same plot is shown |
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irrespective which bioassay set is selected. |
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|
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Currently two control summary curves are generated in one plot; The |
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average intensity of perfect match beads in each assay, and the |
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average intensity of housekeeping beads in each assay. |
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|
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The average intensity I_avg is calculated as |
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I_avg = sum[Iraw_i] / N |
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|
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where N is the number of bead types in the sum, Iraw_i is the raw |
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mean intensity for bead type i. |
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|
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A bead type is considered to belong to the perfect match group if it |
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is annotated with ':pm' in the reporter annotation column '[Rep] |
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Control group id', and a bead type is grouped as housekeeping if it is |
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annotated with 'housekeeping' in reporter annotator column '[Rep] |
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Control group id'. |
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---------------------------------------------------------------------- |
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{{{ |
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Copyright (C) 2009, 2010 Jari Häkkinen |
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This file is part of Illumina plug-in package for BASE. |
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Available at http://baseplugins.thep.lu.se/ |
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BASE main site: http://base.thep.lu.se/ |
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This is free software; you can redistribute it and/or |
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modify it under the terms of the GNU General Public License |
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as published by the Free Software Foundation; either version 3 |
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of the License, or (at your option) any later version. |
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The software is distributed in the hope that it will be useful, |
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but WITHOUT ANY WARRANTY; without even the implied warranty of |
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MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the |
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GNU General Public License for more details. |
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|
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You should have received a copy of the GNU General Public License |
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along with BASE. If not, see <http://www.gnu.org/licenses/>. |
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}}} |
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