Where are they now? **tldr** ![bar](http://eagle.fish.washington.edu/cnidarian/skitch/Screenshot_3_13_15__9_20_AM_1AB34D8C.png) --- In an effort to find out where heat stress induced differentially methylated loci are in the oyster genome (to ultimately inform on function) I have been using `bedtools` to see where the DMLs lie on the genome. As this was done on an array platform I also felt I need to take into consideration where probes were, noting that they were not randomly distributed across the genome but rather targetted to genes. I have determined the proportion of DMLs (n=10028, 10148, 11690) for each oyster that fall within a given genomic feature and compared that to the proporiton of total probes (n=697753) that fall within each genomic feature. For example in just looking at Oyster 2 DMLs and DEGs ... ``` !intersectbed \ -wb \ -a ./data/2014.07.02.colson/genomeBrowserTracks/logFC_HS-preHS/2014.07.02.2M_sig.bedGraph \ -b /Users/sr320/data-genomic/tentacle/Cuffdiff_geneexp.sig.gtf \ | cut -f 6 \ | sort | uniq -c !intersectbed \ -wb \ -a /Users/sr320/git-repos/paper-Temp-stress/ipynb/data/array-design/OID40453_probe_locations.gff \ -b /Users/sr320/data-genomic/tentacle/Cuffdiff_geneexp.sig.gtf \ | cut -f 11 \ | sort | uniq -c ``` output: 880 Cufflinks 117460 Cufflinks ``` # Enter the data comparing Oyster 2 then Probes obs = array([[880, 10028], [117460, 697753]]) # Calculate the chi-square test chi2_corrected = stats.chi2_contingency(obs, correction=True) chi2_uncorrected = stats.chi2_contingency(obs, correction=False) # Print the result print('CHI SQUARE') print('The corrected chi2 value is {0:5.3f}, with p={1:5.3f}'.format(chi2_corrected[0], chi2_corrected[1])) print('The uncorrected chi2 value is {0:5.3f}, with p={1:5.3f}'.format(chi2_uncorrected[0], chi2_uncorrected[1])) ``` output: CHI SQUARE The corrected chi2 value is 352.138, with p=0.000 The uncorrected chi2 value is 352.654, with p=0.000 ~ [jupyter notebook](http://nbviewer.ipython.org/github/sr320/paper-Temp-stress/blob/master/ipynb/Array-feature-overlap-04.ipynb) --- To be honest I feel like I am missing some nuance in the analysis, however at this point I believe I will keep pushing through by seeing of the results break out based on whether the DML is hypo or hypermethylated. If you forgot hear is the breakdown. Oyster | Hypo-methylated | Hyper-methylated | Hypo-3plus-merged | Hypo-3plus-merged --- | --- | --- | --- | --- 2 | 7224 | 2803 | 108 | 4 4 | 6560 | 3587 | 48 | 10 6 | 7645 | 4044 | 53 | 9 This also sheds light on the fact that I am currently ignoring clustering (3-plus), something else to put on the list!