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By choosing proper pairs of catholyte and anolyte, we could achieve desired cathodic and anodic migrating pH gradients in non-IPG-IEF system, effectively eliminating protein precipitation and uncertainty of quantitation existing in routine IEF and 2DE, and enhancing the resolution and sensitivity of IEF.
Proper pairs were extracted using samtools.
Only proper pairs mapping unambiguously to one position were retained.
For downsampling analysis, only reads mapped in proper pairs were considered.
Around 55.4% were mapped as proper pairs while another 13.28% were mapped as singletons.
Proper pairs are defined as reads aligning both within 250 bp of each other and in the correct relative orientation.
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