|
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− | {{STUB}} | + | {{LIVE}} |
| | | |
| {{Vspace}} | | {{Vspace}} |
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| <section begin=abstract /> | | <section begin=abstract /> |
| <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "abstract" --> | | <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "abstract" --> |
− | ...
| + | This unit demonstrates accessing and working with datasets downloaded from NCBI GEO. |
| <section end=abstract /> | | <section end=abstract /> |
| | | |
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| === Objectives === | | === Objectives === |
| <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "objectives" --> | | <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "objectives" --> |
− | ... | + | This unit will ... |
| + | * ... teach downloading and annotating GEO data, and performing differential expression analysis. |
| | | |
| {{Vspace}} | | {{Vspace}} |
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| === Outcomes === | | === Outcomes === |
| <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "outcomes" --> | | <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "outcomes" --> |
− | ... | + | After working through this unit you ... |
| + | * ... can access GEO data; |
| + | * ... are familar with the structure of GEO expression sets; |
| + | * ... can annotate the data, perform differential expression anlysis and critically evaluate the results. |
| | | |
| {{Vspace}} | | {{Vspace}} |
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| === Evaluation === | | === Evaluation === |
| <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "evaluation" --> | | <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "evaluation" --> |
− | <!-- included from "ABC-unit_components.wtxt", section: "eval-TBD" -->
| + | This learning unit can be evaluated for a maximum of 6 marks. If you want to submit the tasks for this unit for credit: |
− | <b>Evaluation: TBD</b><br /> | + | # Create a new page on the student Wiki as a subpage of your User Page. |
− | :This unit can be submitted for evaluation for a maximum of 6 marks. Details TBD.
| + | # There are a number of tasks in which you are explicitly asked you to submit code or other text for credit. Put all of these submission on this one page. |
| + | # When you are done with everything, add the following category tag to the page: |
| + | ::<code><nowiki>[[Category:EVAL-RPR-GEO2R]]</nowiki></code> |
| + | '''Do not''' change your submission page after this tag has been added. The page will be marked and the category tag will be removed by the instructor. |
| | | |
| {{Vspace}} | | {{Vspace}} |
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| <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "contents" --> | | <!-- included from "../components/RPR-GEO2R.components.wtxt", section: "contents" --> |
| | | |
− | | + | {{ABC-unit|RPR-GEO2R.R}} |
− | GEO regex example
| |
− | | |
− | ===Labeling===
| |
− | <div class="mw-collapsible mw-collapsed exercise-box" data-expandtext="Hint" data-collapsetext="Collapse">
| |
− | Write an '''R''' script that creates ''meaningful'' labels for data elements from metadata and shows them in a plot. Use the sample data below - or any other data you are interested in.
| |
− | | |
− | | |
− | | |
− | <div class="mw-collapsible mw-collapsed exercise-box" data-expandtext="Expand" data-collapsetext="Collapse" style="background-color:#EEEEF9;">
| |
− | Sample input data from GEO, and task description ...
| |
− | <div class="mw-collapsible-content">
| |
− | These data were downloaded from the NCBI GEO database using the GEO2R tool, this is a microarray expression data study that compares tumor and metastasis tissue. You can access the dataset [http://www.ncbi.nlm.nih.gov/geo/geo2r/?acc=GSE42952 '''here'''.] Grouping primary PDAC (pancreatic ductal adenocarcinoma) as "tumor" and liver/peritoneal metastasis as "metastasis", an '''R''' script on the server calculates significantly differentially expressed genes using the {{[http://www.bioconductor.org/packages/2.12/bioc/html/limma.html Bioconductor limma package]. I have selected the top 100 genes, and now would like to plot significance (adjusted P value) vs. level of differential expression (logFC). Moreover I would like to vaguely identify the function of each gene if that is discernible from the "Gene title".
| |
− | | |
− | <source lang="text">
| |
− | "ID" "adj.P.Val" "P.Value" "t" "B" "logFC" "Gene.symbol" "Gene.title"
| |
− | "238376_at" "3.69e-19" "4.53e-23" "-49.138515" "42.43328" "-2.202043" "LOC100505564///DEXI" "uncharacterized LOC100505564///Dexi homolog (mouse)"
| |
− | "214041_x_at" "2.36e-17" "8.74e-21" "38.089228" "37.60995" "4.541989" "RPL37A" "ribosomal protein L37a"
| |
− | "241662_x_at" "2.36e-17" "1.03e-20" "-37.793765" "37.45851" "-2.105123" "" ""
| |
− | "231628_s_at" "2.36e-17" "1.16e-20" "-37.574182" "37.34507" "-1.97516" "SERPINB6" "serpin peptidase inhibitor, clade B (ovalbumin), member 6"
| |
− | "224760_at" "3.23e-17" "2.10e-20" "36.500909" "36.77932" "3.798724" "SP1" "Sp1 transcription factor"
| |
− | "214149_s_at" "3.23e-17" "2.38e-20" "36.282193" "36.66167" "4.246787" "ATP6V0E1" "ATPase, H+ transporting, lysosomal 9kDa, V0 subunit e1"
| |
− | "243177_at" "4.15e-17" "3.57e-20" "-35.573827" "36.275" "-1.801709" "" ""
| |
− | "243800_at" "5.63e-17" "5.52e-20" "-34.825113" "35.85663" "-2.018088" "NR1H4" "nuclear receptor subfamily 1, group H, member 4"
| |
− | "238398_s_at" "1.10e-16" "1.21e-19" "-33.519208" "35.10201" "-2.245806" "" ""
| |
− | "1569856_at" "1.48e-16" "1.82e-19" "-32.860752" "34.70891" "-1.810438" "TPP2" "tripeptidyl peptidase II"
| |
− | "1555116_s_at" "1.51e-16" "2.14e-19" "-32.598656" "34.55" "-1.990665" "SLC11A1" "solute carrier family 11 (proton-coupled divalent metal ion transporters), member 1"
| |
− | "218733_at" "1.51e-16" "2.23e-19" "32.535823" "34.51169" "2.764663" "MSL2" "male-specific lethal 2 homolog (Drosophila)"
| |
− | "201225_s_at" "2.72e-16" "4.33e-19" "31.497695" "33.86667" "3.447828" "SRRM1" "serine/arginine repetitive matrix 1"
| |
− | "217052_x_at" "4.45e-16" "7.64e-19" "30.636232" "33.31345" "1.601527" "" ""
| |
− | "1569348_at" "5.24e-16" "9.65e-19" "-30.289176" "33.08577" "-1.793925" "TPTEP1" "transmembrane phosphatase with tensin homology pseudogene 1"
| |
− | "219492_at" "6.96e-16" "1.37e-18" "29.777415" "32.74483" "3.586919" "CHIC2" "cysteine-rich hydrophobic domain 2"
| |
− | "215047_at" "7.51e-16" "1.58e-18" "-29.567379" "32.60307" "-2.033635" "TRIM58" "tripartite motif containing 58"
| |
− | "232877_at" "7.51e-16" "1.66e-18" "-29.491388" "32.55151" "-1.65225" "" ""
| |
− | "229265_at" "7.51e-16" "1.75e-18" "29.419139" "32.50236" "3.933071" "SKI" "v-ski sarcoma viral oncogene homolog (avian)"
| |
− | "1553842_at" "8.16e-16" "2.00e-18" "-29.226409" "32.37061" "-1.832581" "BEND2" "BEN domain containing 2"
| |
− | "220791_x_at" "1.11e-15" "2.87e-18" "-28.71601" "32.01715" "-1.969381" "SCN11A" "sodium channel, voltage-gated, type XI, alpha subunit"
| |
− | "212911_at" "1.17e-15" "3.15e-18" "28.584094" "31.92471" "2.143175" "DNAJC16" "DnaJ (Hsp40) homolog, subfamily C, member 16"
| |
− | "243464_at" "1.22e-15" "3.43e-18" "-28.463254" "31.83963" "-1.675747" "" ""
| |
− | "243823_at" "1.30e-15" "3.81e-18" "-28.316669" "31.7359" "-1.499823" "" ""
| |
− | "201533_at" "1.56e-15" "4.80e-18" "27.999089" "31.5092" "4.054743" "CTNNB1" "catenin (cadherin-associated protein), beta 1, 88kDa"
| |
− | "210878_s_at" "1.59e-15" "5.06e-18" "27.927536" "31.45775" "2.982033" "KDM3B" "lysine (K)-specific demethylase 3B"
| |
− | "227712_at" "3.18e-15" "1.05e-17" "26.938855" "30.73223" "2.426311" "LYRM2" "LYR motif containing 2"
| |
− | "228520_s_at" "3.56e-15" "1.22e-17" "26.742683" "30.58495" "3.744881" "APLP2" "amyloid beta (A4) precursor-like protein 2"
| |
− | "210242_x_at" "3.80e-15" "1.36e-17" "26.605262" "30.48111" "1.815311" "ST20" "suppressor of tumorigenicity 20"
| |
− | "217301_x_at" "3.80e-15" "1.40e-17" "26.565414" "30.45089" "3.275566" "RBBP4" "retinoblastoma binding protein 4"
| |
− | "1557551_at" "6.17e-15" "2.35e-17" "-25.892664" "29.93351" "-1.78824" "" ""
| |
− | "201392_s_at" "6.17e-15" "2.42e-17" "25.856344" "29.90519" "3.283483" "IGF2R" "insulin-like growth factor 2 receptor"
| |
− | "210371_s_at" "7.18e-15" "2.91e-17" "25.62344" "29.72255" "3.463431" "RBBP4" "retinoblastoma binding protein 4"
| |
− | "204252_at" "9.08e-15" "3.79e-17" "25.291186" "29.45902" "2.789842" "CDK2" "cyclin-dependent kinase 2"
| |
− | "243200_at" "1.04e-14" "4.48e-17" "-25.082134" "29.29138" "-1.539093" "" ""
| |
− | "201140_s_at" "1.16e-14" "5.13e-17" "24.916407" "29.15746" "2.834707" "RAB5C" "RAB5C, member RAS oncogene family"
| |
− | "1559066_at" "1.23e-14" "5.57e-17" "-24.813534" "29.07387" "-1.595061" "" ""
| |
− | "201123_s_at" "1.27e-14" "5.91e-17" "24.741268" "29.01494" "4.870779" "EIF5A" "eukaryotic translation initiation factor 5A"
| |
− | "218291_at" "1.41e-14" "6.83e-17" "24.565645" "28.87099" "2.605328" "LAMTOR2" "late endosomal/lysosomal adaptor, MAPK and MTOR activator 2"
| |
− | "217704_x_at" "1.41e-14" "6.91e-17" "-24.550405" "28.85845" "-1.711476" "SUZ12P1" "suppressor of zeste 12 homolog pseudogene 1"
| |
− | "227338_at" "1.44e-14" "7.22e-17" "-24.498114" "28.81536" "-2.927581" "LOC440983" "hypothetical gene supported by BC066916"
| |
− | "210231_x_at" "1.64e-14" "8.47e-17" "24.305184" "28.65556" "4.548338" "SET" "SET nuclear oncogene"
| |
− | "225289_at" "1.86e-14" "9.82e-17" "24.127523" "28.50726" "3.062123" "STAT3" "signal transducer and activator of transcription 3 (acute-phase response factor)"
| |
− | "204658_at" "1.93e-14" "1.04e-16" "24.056703" "28.44783" "2.868797" "TRA2A" "transformer 2 alpha homolog (Drosophila)"
| |
− | "208819_at" "2.54e-14" "1.40e-16" "23.705016" "28.15009" "2.593365" "RAB8A" "RAB8A, member RAS oncogene family"
| |
− | "210011_s_at" "2.58e-14" "1.46e-16" "23.660126" "28.11176" "2.309763" "EWSR1" "EWS RNA-binding protein 1"
| |
− | "202397_at" "2.58e-14" "1.48e-16" "23.638422" "28.0932" "4.332132" "NUTF2" "nuclear transport factor 2"
| |
− | "1552628_a_at" "2.86e-14" "1.68e-16" "23.492249" "27.96778" "2.892763" "HERPUD2" "HERPUD family member 2"
| |
− | "233757_x_at" "3.85e-14" "2.31e-16" "23.123802" "27.64812" "2.430056" "" ""
| |
− | "201545_s_at" "5.07e-14" "3.16e-16" "22.767216" "27.33385" "2.568005" "PABPN1" "poly(A) binding protein, nuclear 1"
| |
− | "1562463_at" "5.07e-14" "3.17e-16" "-22.763883" "27.33089" "-1.119718" "" ""
| |
− | "219859_at" "5.41e-14" "3.45e-16" "-22.669239" "27.24664" "-1.787549" "CLEC4E" "C-type lectin domain family 4, member E"
| |
− | "1569136_at" "6.91e-14" "4.50e-16" "-22.372385" "26.98011" "-1.95396" "MGAT4A" "mannosyl (alpha-1,3-)-glycoprotein beta-1,4-N-acetylglucosaminyltransferase, isozyme A"
| |
− | "208601_s_at" "7.15e-14" "4.74e-16" "-22.314594" "26.92781" "-1.323653" "TUBB1" "tubulin, beta 1 class VI"
| |
− | "226194_at" "1.11e-13" "7.47e-16" "21.813583" "26.46872" "2.331245" "CHAMP1" "chromosome alignment maintaining phosphoprotein 1"
| |
− | "217877_s_at" "1.15e-13" "7.93e-16" "21.748093" "26.40795" "2.862688" "GPBP1L1" "GC-rich promoter binding protein 1-like 1"
| |
− | "225371_at" "1.25e-13" "8.73e-16" "21.644444" "26.31139" "2.518013" "GLE1" "GLE1 RNA export mediator homolog (yeast)"
| |
− | "1563431_x_at" "1.44e-13" "1.02e-15" "21.472848" "26.15053" "1.874743" "CALM3" "calmodulin 3 (phosphorylase kinase, delta)"
| |
− | "211505_s_at" "1.45e-13" "1.06e-15" "21.437744" "26.11746" "2.642609" "STAU1" "staufen double-stranded RNA binding protein 1"
| |
− | "201585_s_at" "1.45e-13" "1.07e-15" "21.430113" "26.11027" "2.787833" "SFPQ" "splicing factor proline/glutamine-rich"
| |
− | "225197_at" "1.75e-13" "1.31e-15" "21.212989" "25.90451" "2.845005" "" ""
| |
− | "220336_s_at" "1.83e-13" "1.41e-15" "-21.132294" "25.82752" "-1.848273" "GP6" "glycoprotein VI (platelet)"
| |
− | "216515_x_at" "1.83e-13" "1.42e-15" "21.128023" "25.82343" "2.877477" "MIR1244-2///MIR1244-3///MIR1244-1///PTMAP5///PTMA" "microRNA 1244-2///microRNA 1244-3///microRNA 1244-1///prothymosin, alpha pseudogene 5///prothymosin, alpha"
| |
− | "241773_at" "3.49e-13" "2.74e-15" "-20.441442" "25.15639" "-1.835223" "" ""
| |
− | "1558011_at" "3.89e-13" "3.15e-15" "-20.297118" "25.01342" "-1.577874" "LOC100510697" "putative POM121-like protein 1-like"
| |
− | "215240_at" "3.89e-13" "3.15e-15" "-20.29699" "25.01329" "-1.613308" "ITGB3" "integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61)"
| |
− | "233746_x_at" "3.95e-13" "3.25e-15" "20.265986" "24.98245" "2.364699" "HYPK///SERF2" "huntingtin interacting protein K///small EDRK-rich factor 2"
| |
− | "1555338_s_at" "4.10e-13" "3.42e-15" "-20.214797" "24.93143" "-1.280803" "AQP10" "aquaporin 10"
| |
− | "217714_x_at" "4.12e-13" "3.48e-15" "20.195128" "24.91179" "2.247023" "STMN1" "stathmin 1"
| |
− | "202276_at" "4.75e-13" "4.08e-15" "20.035595" "24.75183" "2.654202" "SHFM1" "split hand/foot malformation (ectrodactyly) type 1"
| |
− | "225414_at" "6.34e-13" "5.52e-15" "19.733786" "24.44585" "3.287225" "RNF149" "ring finger protein 149"
| |
− | "243930_x_at" "7.43e-13" "6.64e-15" "-19.55046" "24.2578" "-1.219467" "" ""
| |
− | "1569263_at" "7.43e-13" "6.66e-15" "-19.548534" "24.25581" "-1.662363" "" ""
| |
− | "1554876_a_at" "8.55e-13" "7.77e-15" "-19.397142" "24.09923" "-1.388081" "S100Z" "S100 calcium binding protein Z"
| |
− | "220001_at" "1.08e-12" "9.97e-15" "-19.15375" "23.84505" "-1.412727" "PADI4" "peptidyl arginine deiminase, type IV"
| |
− | "228170_at" "1.12e-12" "1.05e-14" "-19.106672" "23.79554" "-1.840114" "OLIG1" "oligodendrocyte transcription factor 1"
| |
− | "211445_x_at" "1.29e-12" "1.22e-14" "-18.959325" "23.63981" "-1.134266" "NACAP1" "nascent-polypeptide-associated complex alpha polypeptide pseudogene 1"
| |
− | "1555311_at" "1.33e-12" "1.27e-14" "-18.91869" "23.59666" "-1.45603" "" ""
| |
− | "201643_x_at" "1.47e-12" "1.43e-14" "18.808994" "23.47974" "1.867155" "KDM3B" "lysine (K)-specific demethylase 3B"
| |
− | "216449_x_at" "1.51e-12" "1.48e-14" "18.773094" "23.44134" "3.178009" "HSP90B1" "heat shock protein 90kDa beta (Grp94), member 1"
| |
− | "218680_x_at" "1.51e-12" "1.50e-14" "18.763896" "23.43149" "2.262739" "HYPK///SERF2" "huntingtin interacting protein K///small EDRK-rich factor 2"
| |
− | "225954_s_at" "1.65e-12" "1.67e-14" "18.662853" "23.32298" "2.405388" "MIDN" "midnolin"
| |
− | "203102_s_at" "1.65e-12" "1.68e-14" "18.658192" "23.31796" "2.476697" "MGAT2" "mannosyl (alpha-1,6-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase"
| |
− | "1569345_at" "1.69e-12" "1.74e-14" "18.624203" "23.28133" "1.236884" "" ""
| |
− | "214001_x_at" "1.71e-12" "1.78e-14" "18.598496" "23.25358" "2.570012" "" ""
| |
− | "231812_x_at" "1.72e-12" "1.81e-14" "18.583236" "23.2371" "1.678685" "PHAX" "phosphorylated adaptor for RNA export"
| |
− | "232075_at" "1.93e-12" "2.06e-14" "-18.462717" "23.10643" "-2.150701" "WDR61" "WD repeat domain 61"
| |
− | "200669_s_at" "1.96e-12" "2.12e-14" "18.438729" "23.08033" "1.891968" "UBE2D3" "ubiquitin-conjugating enzyme E2D 3"
| |
− | "236995_x_at" "2.04e-12" "2.23e-14" "-18.389604" "23.02677" "-1.879369" "TFEC" "transcription factor EC"
| |
− | "218008_at" "2.24e-12" "2.48e-14" "18.291537" "22.91946" "2.445428" "TMEM248" "transmembrane protein 248"
| |
− | "217140_s_at" "2.30e-12" "2.56e-14" "18.260017" "22.88485" "3.983721" "VDAC1" "voltage-dependent anion channel 1"
| |
− | "210183_x_at" "2.46e-12" "2.79e-14" "18.183339" "22.80044" "1.79105" "PNN" "pinin, desmosome associated protein"
| |
− | "216954_x_at" "2.46e-12" "2.80e-14" "-18.177967" "22.79451" "-1.090193" "ATP5O" "ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit"
| |
− | "207688_s_at" "2.53e-12" "2.92e-14" "18.141153" "22.75385" "2.492309" "INHBC" "inhibin, beta C"
| |
− | "218020_s_at" "2.63e-12" "3.06e-14" "18.095669" "22.70351" "1.772689" "ZFAND3" "zinc finger, AN1-type domain 3"
| |
− | "217756_x_at" "3.12e-12" "3.67e-14" "17.930201" "22.51939" "1.914366" "SERF2" "small EDRK-rich factor 2"
| |
− | "214150_x_at" "3.42e-12" "4.07e-14" "-17.835551" "22.41336" "-1.177963" "ATP6V0E1" "ATPase, H+ transporting, lysosomal 9kDa, V0 subunit e1"
| |
− | "208750_s_at" "3.48e-12" "4.18e-14" "17.812279" "22.38721" "2.649599" "ARF1" "ADP-ribosylation factor 1"
| |
− | "201749_at" "3.59e-12" "4.42e-14" "17.761415" "22.32994" "1.917794" "ECE1" "endothelin converting enzyme 1"
| |
− | </source>
| |
− | </div>
| |
− | </div>
| |
− | | |
− | | |
− | | |
− | | |
− | <div class="mw-collapsible-content exercise-box">
| |
− | <div class="mw-collapsible mw-collapsed" data-expandtext="Solution" data-collapsetext="Collapse">
| |
− | Read the data into '''R'''. Plot log(P) against log(FC). Define some regular expressions that identify keywords in the gene title: things like "X-ase", "Y factor", "Z gene" etc. Apply these to the gene titles using {{R|regex()||regexpr()}} and store the results by applying {{R|regmatches()}} to the text. Then use {{R|graphics|text()}} to plot the extracted strings.
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− | | |
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− | <div class="mw-collapsible-content exercise-box">
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− | <source lang="R">
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− | #GEO-hits.R
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− | # bs - Sept. 2013
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− | dat <- read.table("GEO-hits_100.txt", header = TRUE) # this is a file of GEO
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− | # differential expression data
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− | head(dat)
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− | plot(-log(dat[,"adj.P.Val"]), dat[,"logFC"], cex=0.7, pch=16, col="#BB0000")
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− | # Note that all these genes have at least one log of
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− | # differential expression - up or down. As a trend,
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− | # higher probabilities are found for higher levels of
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− | # differential expression.
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− | # The dataframe produced by R's read.table() function
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− | # defines all character-containing rows as _factors_.
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− | # However to process them as strings, we need to convert
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− | # them to characters.
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− | dat[,"Gene.title"] <- as.character(dat[,"Gene.title"])
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− | | |
− | # First, let's define some regexes for keywords to guess
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− | # a function ...
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− | # (Note the need for doubled escape characters in R!)
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− | r <- c( "\\b(\\w+ase)\\b") # peptidase, kinase ...
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− | r <- c(r, "\\b(?!factor)(\\w+or)") # suppressor, adaptor ...
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− | r <- c(r, "\\b(\\w+)\\b\\s(factor|protein|homolog)") # the preceeding word ...
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− | # Now iterate over the Gene.title column and for each row try all regular
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− | # expressions.
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− | for (i in 1:nrow(dat)) { # for all rows ...
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− | for (j in 1:length(r)) { # for all regular expressions
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− | dat[i,"Function.guess"] <- "" # clear the contents of the column
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− | M <- regexpr(r[j], dat[i, "Gene.title"], perl = TRUE)
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− | if (M[1] > 0) {
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− | dat[i,"Function.guess"] <- regmatches(dat[i,"Gene.title"], M)
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− | break # stop regexing if something was found
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− | }
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− | }
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− | }
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− | dat[,"Function.guess"] # check what we found ...
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− | # ... and plot the strings to the right of its point.
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− | text(-log(dat[,"adj.P.Val"]), dat[,"logFC"], dat[,"Function.guess"], cex=0.4, pos=4)
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− | # I'm not sure we are actually learning anything important from this.
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− | # But the code was merely meant to illustrate how
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− | # to work with regular expressions in R (and introduce you to GEO
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− | # differential expression data on the side). Mission accomplished.
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− | </source>
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Line 345: |
Line 165: |
| :2017-08-05 | | :2017-08-05 |
| <b>Modified:</b><br /> | | <b>Modified:</b><br /> |
− | :2017-08-05 | + | :2017-11-11 |
| <b>Version:</b><br /> | | <b>Version:</b><br /> |
− | :0.1 | + | :1.0 |
| <b>Version history:</b><br /> | | <b>Version history:</b><br /> |
| + | *1.0 First live version |
| *0.1 First stub | | *0.1 First stub |
| </div> | | </div> |