Rna004

README.md

@@ -78,6 +78,8 @@ By default, DNA PromethION reads (R9.4.1) will be simulated. Specify the `-x STR
 - `rna-r9-prom`: direct RNA on PromethION R9.4.1 flowcells
 - `dna-r10-min`: genomic DNA on MinION R10.4.1 flowcells
 - `dna-r10-prom`: genomic DNA on PromethION R10.4.1 flowcells
+- `rna004-min`: direct RNA on MinION RNA004 flowcells
+- `rna004-prom`: direct RNA on promethION RNA004 flowcells
 
 If a genomic DNA profile is selected, the input reference must be the **reference genome in *FASTA* format**. *squigulator* will randomly sample the genome from a uniform distribution and generate reads whose lengths are from a gamma distribution (based on `-r`). If a direct RNA profile is selected, the input reference must be the **transcriptome in *FASTA* format**. For RNA, *squigulator* will randomly pick transcripts from a uniform distribution and the whole transcript length is simulated.
 

docs/man.md

@@ -3,7 +3,7 @@
 Basic options in *squigulator* are as below:
 
 - `-o FILE`: SLOW5/BLOW5 file to write.
-- `-x STR`: Parameter profile (always applied before other options). Available profiles are: *dna-r9-min*, *dna-r9-prom, rna-r9-min*, *rna-r9-prom*, *dna-r10-min*, *dna-r10-prom*. [default: dna-r9-prom]
+- `-x STR`: Parameter profile (always applied before other options). Available profiles are: *dna-r9-min*, *dna-r9-prom, rna-r9-min*, *rna-r9-prom*, *dna-r10-min*, *dna-r10-prom*, *rna004-min, *rna004-prom* [default: dna-r9-prom]
 - `-n INT`: Number of reads to simulate. [default: 4000]
 - `-r INT `: Mean read length (estimated mean only, unused for RNA) [default: 10000]
 - `-f INT`:  fold coverage to simulate (incompatible with -n)
@@ -37,4 +37,5 @@ Developer options (which are not much tested and error handling) are as below:
 -  `--range FLOAT`:              ADC range (see [here](https://hasindu2008.github.io/slow5specs/summary))
 -  `--offset-mean FLOAT`:        ADC offset mean (see [here](https://hasindu2008.github.io/slow5specs/summary))
 -  `--offset-std FLOAT`:         ADC offset standard deviation (see [here](https://hasindu2008.github.io/slow5specs/summary))
-
+-  `--median-before-mean`:      Median before mean (see [here](https://hasindu2008.github.io/slow5specs/summary))
+-  `--median-before-std`:       Median before standard deviation (see [here](https://hasindu2008.github.io/slow5specs/summary))

docs/profile.md

@@ -0,0 +1,98 @@
+# Parameter profiles
+
+-x option. Sets the parameters below.
+
+## R9 profiles
+
+- dna-r9-min: DNA R9.4.1 MinION (or GridION)
+- dna-r9-prom: DNA R9.4.1 promethION (or P2solo)
+- rna-r9-min: RNA R9.4.1 MinION (or GridION)
+- rna-r9-prom:  RNA R9.4.1 promethION (or P2solo)
+
+| Profile              | dna-r9-min   | dna-r9-prom   | rna-r9-min   | rna-r9-prom   |
+|----------------------|--------------|---------------|--------------|---------------|
+| digitisation         | 8192         | 2048          | 8192         | 2048          |
+| sample-rate          | 4000         | 4000          | 3012         | 3000          |
+| bps                  | 450          | 450           | 70           | 70            |
+| range                | 1443.030273  | 748.5801      | 1126.47      | 548.788269    |
+| offset-mean          | 13.7222605   | -237.4102     | 4.65491888   | -231.9440589  |
+| Offset STD           | 10.25279688  | 14.1575       | 4.115262472  | 12.87185278   |
+| Median Before Mean   | 200.815801   | 214.2890337   | 242.6584118  | 238.5286796   |
+| Median Before STD    | 20.48933762  | 18.0127916    | 10.60230888  | 21.1871794    |
+| Dwell Mean           | 9.0          | 9.0           | 43.0         | 43.0          |
+| Dwell STD            | 4.0          | 4.0           | 35.0         | 35.0          |
+
+
+## R10 and RNA004 profiles
+
+- dna-r10-min:  DNA R10.4.1 MinION (or GridION)
+- dna-r10-prom: DNA R10.4.1 promethION (or P2solo)
+- rna004-min: RNA004 MinION (or GridION)
+- rna004-prom: RNA004 promethION (or P2solo)
+
+| Profile             | dna-r10-min   | dna-r10-prom   | rna004-min   | rna004-prom   |
+|---------------------|---------------|-----------------|--------------|---------------|
+| digitisation        | 8192          | 2048            | 2048         | 2048          |
+| sample-rate         | 4000          | 4000            | 4000         | 4000          |
+| bps                 | 400           | 400             | 130          | 130           |
+| range               | 1536.598389   | 281.345551      | TBD  | 299.432068    |
+| offset-mean         | 13.380569389  | -127.5655735    | TBD5 | -259.421128  |
+| offset-std         | 16.311471649  | 19.377283387665 | TBD | 16.010841823643 |
+| median-before-mean  | 202.154074388 | 189.87607393756 | TBD | 189.87607393756 |
+| median-before-std   | 13.406139242  | 15.788097978713 | TBD | 15.788097978713 |
+| dwell-mean          | 10.0          | 10.0            | TBD         | 31.0          |
+| dwell-std           | 4.0           | 4.0             | TBD          | 4.0           |
+
+## Determining parameters for a profile
+
+Assume S/BLOW5. Need slow5tools and datamash.
+Convert using X and Y. Following methods:
+Assume you have a pore model.
+
+
+- digitisation. This is the [digitisation field in the BLOW5 file](https://hasindu2008.github.io/slow5specs/summary). Observe that this is the same across the whole dataset (Infact, MinIONs/GridIONs so far has 8192 and promethION/P2 has 2048).
+
+  Example command (you should only see one value):
+  ```
+  slow5tools skim -t40 PNXRXX240011_reads_500k.blow5 | cut -f 3 | tail -n+2 | sort -u
+  2048
+  ```
+
+- sample-rate. This is the [sample-rate field in the BLOW5 file](https://hasindu2008.github.io/slow5specs/summary). Observe that this is the same across the whole dataset.
+
+  Example command (you should only see one value):
+  ```
+  slow5tools skim -t40 PNXRXX240011_reads_500k.blow5 | cut -f 6 | tail -n+2 | sort -u
+  4000
+  ```
+
+- bps. This is the translocation speed which can be found on the relevant Guppy/Dorado model. For example, the Dorado model for rna004 is `[email protected]`. The bps is 130.
+
+- range. This is the [digitisation field in the BLOW5 file](https://hasindu2008.github.io/slow5specs/summary).
+
+  Example command (you should only see one value):
+  ```
+  slow5tools skim -t40 PNXRXX240011_reads_500k.blow5 | cut -f 5 | tail -n+2 |  sort -u
+  299.432068
+  ```
+
+- offset-mean and offset-std. This is the mean and standard deviation of the [offset field in the BLOW5 file](https://hasindu2008.github.io/slow5specs/summary).
+
+  Example command to get the two parameters:
+  ```
+   slow5tools skim -t40 PNXRXX240011_reads_500k.blow5 | cut -f 4 | tail -n+2 |  datamash mean 1 sstdev 1
+   -259.421128     16.010841823643
+  ```
+
+-
+- median-before-mean and median-before-std. This is the mean and standard deviation of the [median_before in the BLOW5 file](https://hasindu2008.github.io/slow5specs/summary).
+
+Example command to get the two parameters:
+ ```
+  slow5tools skim -t40 PNXRXX240011_reads_500k.blow5 | awk -v c="median_before" 'NR==1{for (i=1; i<=NF; i++) if ($i==c){p=i; break};} {if ($p!=".") print $p}' | tail -n+2 | datamash mean 1 sstdev 1
+  205.63935594369 8.3994882799157
+ ```
+
+- dwell-mean. This must be equal to the sample_rate/bps, and acts as a sanity check currently.
+
+- dwell-std.

src/model.c

@@ -169,8 +169,12 @@ uint32_t set_model(model_t* model, uint32_t model_id) {
         num_kmer=262144;
         inbuilt_model=r10_4_nucleotide_9mer_template_model_builtin_data;
         assert(num_kmer == (uint32_t)(1 << 2*kmer_size)); //num_kmer should be 4^kmer_size
-    }
-    else{
+    } else if(model_id==MODEL_ID_RNA_RNA004_NUCLEOTIDE){
+        kmer_size=9;
+        num_kmer=262144;
+        inbuilt_model=rna004_130bps_u_to_t_rna_9mer_template_model_builtin_data;
+        assert(num_kmer == (uint32_t)(1 << 2*kmer_size)); //num_kmer should be 4^kmer_size
+    } else{
         assert(0);
     }