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Extract k‐mers specific to germ‐line restricted chromosomes
Now, that we know how to make the 2d k-mer plot, let's find chromosome-specific k-mers. As no one has actually seen the tse-tse data, we can't write here what threshold we should use, that's something we will need to discuss together dependent on the results of the previous section.
determining thresholds
To show you how the thresholds can be terminted, here we show an example of Bradysia tilicola from Hodson et al. 2021. In this preprint, germ-line restricted chromosomes were isolated by comparing a library made of testes and heads of flies. The karyotypes in the two types of tissues, as well as 2d k-mer spectra, are in the figure 1 from the manuscript
Hence for the isolation of 27-mers corresponding to X, GRC or autosomes respectively we chose the following thresholds (boxed on the image)
A
- 125 < head < 175
- 80 < testes < 140
X
- 50 < head < 100
- 60 < testes < 100
GRC
- head < 5
- 15 < testes # this might be too stringent
Introduction
k-mer spectra analysis
- 📖 Introduction to K-mer spectra analysis
- 📖 Basics of genome modeling
- ⚒ manual model fitting (for better understanding of the underlying model)
- ⚒ simple diploid
- ⚒ demonstrating the effect of sequencing error rate on k-mer coverage
- 📖 Common difficulties in characterisation of diploid genomes using k mer spectra analysis
- ⚒ low coverage (pitfall) - to be merged
- ⚒ very homozygous diploid
- ⚒ highly heterozygous diploid
- ⚒ Genome size of a repetitive genome (pitfall)
- ⚒ Wrong ploidy (pitfall)
- 📖 Characterization of polyploid genomes using k mer spectra analysis
- ⚒ Autotetraploid
- ⚒ Allotetraploid
- ⚒ Estimating ploidy (smudgeplot)
- 📖 Genome modeling as a quality control
- ⚒ Contamination (pitfall)
- ⚒ k-mers in an assembly (Mercury/KAT)
- 📖 Analysing genome skimming data
Separation of chromosomes
- 📖Separate sub-genomes of an allopolyploid
- 📖Separating chromosomes by comparison of sequencing libraries
Species assignment using short k-mers
Others