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For instance, let's say that my reads per gene contains the following information for these two transcripts (Reads per gene file obtained with STAR):
**ENST00000361204** 0 0 **0**
**ENST00000255784** 842 0 **842**
So 0 reads per ENST00000361204 transcript and 842 reads per ENST00000255784 transcript.
And the file generated during annotation step contains lines like the following:
chr22 42276719 42290941 circular_RNA/19 0 + 42276719 42276719 0 0 0 4 277 170 169 118 0 4126 12401 14104 19 circRNA SREBF2 **ENST00000361204** 10 11 12 13 chr22:42274127-42276719|chr22:42290941-42293055
chr22 42204878 42206295 circular_RNA/260 0 + 42204878 42204878 0 0 0 3 119 122 85 0 1004 1332 260 circRNA CCDC134 **ENST00000255784** 2 3 4 chr22:42196770-42204878|chr22:42206295-42209267
chr22 42204878 42209826 circular_RNA/81 0 + 42204878 42204878 0 0 0 5 119 122 85 182 72 0 1004 1332 4389 4876 81 circRNA CCDC134 **ENST00000255784** 2 3 4 5 6 chr22:42196770-42204878|chr22:42209826-42221701
Questions for you:
- It seems that a circRNA had 0 reads yet it was annotated (ENST00000361204). Why? What does it mean?
- For ENST00000255784 transcript, it is annotated twice as two different circRNAs. How would I know the weight of this transcript? How sure am I that it would not be a 3rd transcript? Should I use bedfiles? At which step?
- After the annotation step, how do you actually cross the information between the circRNAs that were aligned and annotated? How do you extract the exact coordinates from the reads per gene file?
Thanks a lot for your help.
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