To take your first steps with HiCognition, you can download example data from our dropbox location. The files located there are centered on analysis connected to sister chromatid sensitive Hi-C.
Needed data:
G2_tads_w_size.bed
| The location of TADs in G2 synchronized HeLa cellsG2.fc_1_2_3_4.wOldG2.cis.1000.mcool
| Cis-sister contacts in G2 synchronized HeLa cellsG2.fc_1_2_3_4.wOldG2.trans.1000.mcool
| Trans-sister contacts in G2 synchronized HeLa cellsTo upload the files, follow our upload instructions for genomic regions and genomic features. The metadata settings are the following:
TADs:
Perturbation
| noneCell cycle stage
| G2Genome assembly
| hg19Sizetype
| IntervalValueType
| DerivedMethod
| HiCCooler files:
Perturbation
| noneCell cycle stage
| G2Genome assembly
| hg19ValueType
| InteractionMethod
| HiCNormalization
| ICCFThe .mcool files are quite big, and you should expect them to take a few minutes to upload (depending on your internet connection).
Once the files are uploaded, start preprocessing the two 2D features at TAD-boundaries by following our preprocessing guide. Once preprocessing is finished, you can create a widget collection by clicking on the at the bottom right of the HiCognition start page. This will create an empty widget collection:
By clicking on the region-controls, you can select your TADs as a region. Once that is done, hover over the center +-icon to reveal the widget-type menu:
Once you select the 2D-average widget, you can select one of the two 2D features you added. For example, if you select the cis-sister contacts, your widget collection should look like this:
This picture represents the average behavior of cis-sister contacts around TADs. You can then resize your widget collection to add trans-sister contacts as a second widget. The resulting collection should look like this:
This view now represents the average behavior of cis-sister and trans-sister contacts around TAD-boundaries. If you want to look at the heterogeneity of, e.g., trans-sister contacts around TADs, you can expand the size of your widget collection to add a new slot and fill it with a 2D-embedding widget:
Once you have added the widget, you can select the trans-sister contacts as features to display. Your widget collection should now look like this:
The 2D-embedding widget now allows you to look at the heterogeneity of trans-sister contacts at TADs (see the section about this widget for more details).