Fragment library types
RNA-seq library preparation protocols differ in ways that matter for
quantification: reads can be single-end or paired-end, the protocol can be
unstranded or strand-specific, and paired-end reads have a relative orientation.
salmon describes all of these with a short library-type string passed to
-l/--libType.
The library-type string has up to three parts: relative orientation, strandedness, and directionality.
1. Relative orientation (paired-end only)
Section titled “1. Relative orientation (paired-end only)”| Code | Meaning |
|---|---|
I | Inward — mates point toward each other |
O | Outward — mates point away from each other |
M | Matching — mates point the same way |
2. Strandedness
Section titled “2. Strandedness”| Code | Meaning |
|---|---|
S | Stranded (strand-specific protocol) |
U | Unstranded |
If the protocol is unstranded (U), the string ends here.
3. Directionality (stranded protocols only)
Section titled “3. Directionality (stranded protocols only)”| Code | Meaning |
|---|---|
F | Read 1 (or the single-end read) comes from the forward strand |
R | Read 1 (or the single-end read) comes from the reverse strand |
Examples
Section titled “Examples”| String | Interpretation |
|---|---|
IU | Unstranded paired-end, mates face each other |
ISF | Stranded paired-end, inward, read 1 from the forward strand |
OSR | Stranded paired-end, outward, read 1 from the reverse strand |
U | Unstranded single-end |
SF | Stranded single-end, reads from the forward strand |
SR | Stranded single-end, reads from the reverse strand |
Single-end libraries omit the orientation part, so the only valid single-end
strings are U, SF, and SR.
Correspondence to TopHat library types
Section titled “Correspondence to TopHat library types”| TopHat | salmon (paired-end) | salmon (single-end) |
|---|---|---|
-fr-unstranded | -l IU | -l U |
-fr-firststrand | -l ISR | -l SR |
-fr-secondstrand | -l ISF | -l SF |
The remaining salmon library strings have no direct TopHat equivalent.