levseq 1.2.5__tar.gz → 1.2.6__tar.gz

{levseq-1.2.5/levseq.egg-info → levseq-1.2.6}/PKG-INFO

@@ -1,6 +1,6 @@
 Metadata-Version: 2.1
 Name: levseq
-Version: 1.2.5
+Version: 1.2.6
 Home-page: https://github.com/fhalab/levseq/
 Author: Yueming Long, Emreay Gursoy, Ariane Mora, Francesca-Zhoufan Li
 Author-email: ylong@caltech.edu
@@ -80,7 +80,7 @@ and `minimap2` installed on your path. However, if you have issues we recommend
 We recommend using terminal and a conda environment for installation:
 
 ```
-conda create --name levseq python=3.10 -y
+conda create --name levseq python=3.12 -y
 ```
 
 ```
@@ -93,7 +93,7 @@ conda activate levseq
 ```
 conda install -c bioconda -c conda-forge samtools
 ```
-or for mac users you can use: `brew install samtools`
+
 
 2. Minimap2: https://github.com/lh3/minimap2
 
@@ -110,11 +110,46 @@ operating system (https://docs.docker.com/engine/install/).
 ```
 levseq <name of the run you can make this whatever> <location to data folder> <location of reference csv file>
 ```
+
 #### Run via docker
+If using linux system
+```
+docker pull yueminglong/levseq:levseq-1.2.5-x86
+```
+If using Mac M chips (image tested on M1, M3, and M4)
+```
+docker pull yueminglong/levseq:levseq-1.2.5-arm64
 ```
-docker run --rm -v "$(pwd):/levseq_results" levseq <name> <location to data folder> <location of reference csv file>
+
+```
+docker run --rm -v "$(pwd):/levseq_results" yueminglong/levseq:levseq-1.2.5-<architecture> <name> <location to data folder> <location of reference csv file>
 ```
+Explanation:
+
+--rm: Automatically removes the container after the command finishes.
+
+-v "$(pwd):/levseq\_results": Mounts the current directory ($(pwd)) to /levseq\_results inside the container, ensuring the results are saved to your current directory.
+
+yueminglong/levseq:levseq-1.2.5-\<architecture\>: Specifies the Docker image to run. Replace \<architecture\> with the appropriate platform (e.g., x86).
+
+\<name\>: The name or identifier for the analysis.
+
+\<location to data folder\>: Path to the folder containing input data.
+
+\<location of reference csv file\>: Path to the reference .csv file.
+
+Important Notes:
+
+If the current directory is mounted to the container (via -v "$(pwd):/levseq\_results"), the basecalled result in FASTQ format and the ref.csv file must be located in the current directory.
+
+If these files are not present in the current directory, they will not be processed by the tool.
+
+Output:
+
+The results of the analysis will be saved to your current working directory.
+
 See the [manuscrtipt notebook](https://github.com/fhalab/LevSeq/blob/main/manuscript/notebooks/epPCR_10plates.ipynb) for an example.
+*Note: if using docker, the html and csv final output will be saved in the directory that you are running from instead of in the Platemaps or Results subfolder.
 
 #### Required Arguments
 1. Name of the experiment, this will be the name of the output folder

{levseq-1.2.5 → levseq-1.2.6}/README.md

@@ -33,7 +33,7 @@ and `minimap2` installed on your path. However, if you have issues we recommend
 We recommend using terminal and a conda environment for installation:
 
 ```
-conda create --name levseq python=3.10 -y
+conda create --name levseq python=3.12 -y
 ```
 
 ```
@@ -46,7 +46,7 @@ conda activate levseq
 ```
 conda install -c bioconda -c conda-forge samtools
 ```
-or for mac users you can use: `brew install samtools`
+
 
 2. Minimap2: https://github.com/lh3/minimap2
 
@@ -63,11 +63,46 @@ operating system (https://docs.docker.com/engine/install/).
 ```
 levseq <name of the run you can make this whatever> <location to data folder> <location of reference csv file>
 ```
+
 #### Run via docker
+If using linux system
+```
+docker pull yueminglong/levseq:levseq-1.2.5-x86
+```
+If using Mac M chips (image tested on M1, M3, and M4)
+```
+docker pull yueminglong/levseq:levseq-1.2.5-arm64
 ```
-docker run --rm -v "$(pwd):/levseq_results" levseq <name> <location to data folder> <location of reference csv file>
+
+```
+docker run --rm -v "$(pwd):/levseq_results" yueminglong/levseq:levseq-1.2.5-<architecture> <name> <location to data folder> <location of reference csv file>
 ```
+Explanation:
+
+--rm: Automatically removes the container after the command finishes.
+
+-v "$(pwd):/levseq\_results": Mounts the current directory ($(pwd)) to /levseq\_results inside the container, ensuring the results are saved to your current directory.
+
+yueminglong/levseq:levseq-1.2.5-\<architecture\>: Specifies the Docker image to run. Replace \<architecture\> with the appropriate platform (e.g., x86).
+
+\<name\>: The name or identifier for the analysis.
+
+\<location to data folder\>: Path to the folder containing input data.
+
+\<location of reference csv file\>: Path to the reference .csv file.
+
+Important Notes:
+
+If the current directory is mounted to the container (via -v "$(pwd):/levseq\_results"), the basecalled result in FASTQ format and the ref.csv file must be located in the current directory.
+
+If these files are not present in the current directory, they will not be processed by the tool.
+
+Output:
+
+The results of the analysis will be saved to your current working directory.
+
 See the [manuscrtipt notebook](https://github.com/fhalab/LevSeq/blob/main/manuscript/notebooks/epPCR_10plates.ipynb) for an example.
+*Note: if using docker, the html and csv final output will be saved in the directory that you are running from instead of in the Platemaps or Results subfolder.
 
 #### Required Arguments
 1. Name of the experiment, this will be the name of the output folder
@@ -89,4 +124,4 @@ For more details or trouble shooting please look at our [computational_protocols
 
 #### Citing
 
-If you have found LevSeq useful, please cite out [paper](https://doi.org/10.1101/2024.09.04.611255).
+If you have found LevSeq useful, please cite out [paper](https://doi.org/10.1101/2024.09.04.611255).

{levseq-1.2.5 → levseq-1.2.6}/levseq/__init__.py

@@ -18,7 +18,7 @@
 __title__ = 'levseq'
 __description__ = 'LevSeq nanopore sequencing'
 __url__ = 'https://github.com/fhalab/levseq/'
-__version__ = '1.2.5'
+__version__ = '1.2.6'
 __author__ = 'Yueming Long, Emreay Gursoy, Ariane Mora, Francesca-Zhoufan Li'
 __author_email__ = 'ylong@caltech.edu'
 __license__ = 'GPL3'

levseq-1.2.6/levseq/coordinates.py

@@ -0,0 +1,76 @@
+import esm
+import torch
+import pandas as pd
+from sklearn.decomposition import PCA
+import os
+import argparse
+
+def preprocess_sequence(sequence):
+    """
+    Preprocesses the amino acid sequence by removing everything after the first '*' (stop codon).
+    """
+    if '*' in sequence:
+        sequence = sequence.split('*')[0]  # Take everything before the first '*'
+    return sequence
+
+def process_file(input_file, output_file=None):
+    # Load the dataset
+    data = pd.read_csv(input_file)
+
+    # Remove the "Unnamed: 0" column if it exists
+    if 'Unnamed: 0' in data.columns:
+        data = data.drop(columns=['Unnamed: 0'])
+
+    # Create the ID column as the combination of `Plate` and `Well`
+    data['ID'] = data['Plate'] + '-' + data['Well']
+    data = data[['ID'] + [col for col in data.columns if col != 'ID']]  # Reorder to make ID the first column
+
+    # Filter valid sequences from the `aa_sequence` column
+    valid_sequences = data['aa_sequence'].dropna()
+    valid_sequences = valid_sequences[~valid_sequences.str.contains('#N.A.#|Deletion')]
+
+    # Preprocess sequences to handle stop codons
+    valid_sequences = valid_sequences.apply(preprocess_sequence)
+
+    # Load the ESM-2 model
+    model, alphabet = esm.pretrained.esm2_t33_650M_UR50D()
+    batch_converter = alphabet.get_batch_converter()
+
+    # Prepare sequences for embedding
+    sequences = valid_sequences.tolist()
+    sequence_names = [f"Sequence {i}" for i in range(len(sequences))]
+    batch_labels, batch_strs, batch_tokens = batch_converter(list(zip(sequence_names, sequences)))
+
+    # Extract embeddings
+    with torch.no_grad():
+        results = model(batch_tokens, repr_layers=[33])
+        embeddings = results["representations"][33]  # Use the top (last) layer representations
+
+    # Average embeddings across residues for sequence-level representation
+    sequence_embeddings = embeddings.mean(1).numpy()
+
+    # Dimensionality Reduction using PCA
+    pca = PCA(n_components=2)
+    xy_coordinates = pca.fit_transform(sequence_embeddings)
+
+    # Add x, y coordinates back to the dataframe
+    xy_df = pd.DataFrame(xy_coordinates, columns=['x_coordinate', 'y_coordinate'], index=valid_sequences.index)
+    data = pd.concat([data, xy_df], axis=1)
+
+    # Determine output file location
+    if output_file is None:
+        input_name, input_ext = os.path.splitext(input_file)
+        output_file = f"{input_name}_xy{input_ext}"
+
+    # Save the updated dataframe to a file
+    data.to_csv(output_file, index=False)
+    print(f"Processed data with x, y coordinates saved to: {output_file}")
+
+if __name__ == "__main__":
+    parser = argparse.ArgumentParser(description="Generate x, y coordinates for amino acid sequences")
+    parser.add_argument('input_file', type=str, help="Path to the input CSV file")
+    parser.add_argument('--output_file', type=str, default=None, help="Path to save the output CSV file (optional)")
+    args = parser.parse_args()
+
+    process_file(args.input_file, args.output_file)
+

{levseq-1.2.5 → levseq-1.2.6}/levseq/run_levseq.py

@@ -321,30 +321,36 @@ def create_df_v(variants_df):
     df_variants_["Plate"] = df_variants_["Plate"].apply(
         lambda x: f"0{x}" if len(x) == 1 else x
     )
-    # Rename columns as per the request
+
+    # First rename columns as before
     df_variants_.rename(columns={
         "Variant": "nucleotide_mutation",
-        "Substitutions": "amino-acid_substitutions",
+        "Substitutions": "amino_acid_substitutions",
         "nc_variant": "nt_sequence",
         "aa_variant": "aa_sequence"
-	    },inplace=True)
-
-	# Select the desired columns in the desired order
-    restructured_df = df_variants_[[
-            "barcode_plate",
-            "Plate",
-            "Well",
-            "Alignment Count",
-            "nucleotide_mutation",
-            "amino-acid_substitutions",
-            "Alignment Probability",
-            "Average mutation frequency",
-            "P value",
-            "P adj. value",
-            "nt_sequence",
-            "aa_sequence",
-        ]
-    ]
+        }, inplace=True)
+
+    # Create a copy for restructuring to avoid affecting the original
+    restructured_df = df_variants_.copy()
+    restructured_df.columns = restructured_df.columns.str.lower().str.replace('[\s-]', '_', regex=True)
+    # Fix the specific column name
+    restructured_df.columns = restructured_df.columns.str.replace('p_adj._value', 'p_adj_value')
+
+    # Select the desired columns in the desired order
+    restructured_df = restructured_df[[
+        "barcode_plate",
+        "plate",
+        "well",
+        "alignment_count",
+        "nucleotide_mutation",
+        "amino_acid_substitutions",
+        "alignment_probability",
+        "average_mutation_frequency",
+        "p_value",
+        "p_adj_value",
+        "nt_sequence",
+        "aa_sequence"
+    ]]
 
     return restructured_df, df_variants_