diff --git a/Chiron+weight_visual/README.md b/Chiron+weight_visual/README.md index 193205f..585ae84 100644 --- a/Chiron+weight_visual/README.md +++ b/Chiron+weight_visual/README.md @@ -1,174 +1,2 @@ -# Chiron -## A basecaller for Oxford Nanopore Technologies' sequencers -Using a deep learning CNN+RNN+CTC structure to establish end-to-end basecalling for the nanopore sequencer. -Built with **TensorFlow** and python 2.7. - -If you found Chiron useful, please consider to cite: -> Teng, H., et al. (2017). Chiron: Translating nanopore raw signal directly into nucleotide sequence using deep learning. [bioRxiv 179531] (https://www.biorxiv.org/content/early/2017/09/12/179531) - ---- - -## Install -### Install using `pip` (recommended) -If you currently have TensorFlow installed on your system, we would advise you to create a virtual environment to install Chiron into, this way there is no clash of versions etc. - -If you would like to do this, the best options would be [`virtualenv`](https://virtualenv.pypa.io/en/stable/installation/), the more user-friendly [`virtualenvwrapper`](https://virtualenvwrapper.readthedocs.io/en/latest/install.html), or through [anaconda](https://docs.continuum.io/anaconda/install/). After installing one of these and activating the virtual environment you will be installing Chiron into, continue with the rest of the installation instructions as normal. - -To install with `pip`: - -``` -pip install chiron -``` -This will install Chiron, the CPU-only distribution of TensorFlow (and it's dependencies), and [`h5py`](https://github.com/h5py/h5py) (required for reading in `.fast5` files). - -**Note**: If you are after the GPU version, follow the steps in the following section. - -### Install from GitHub -This is currently the best install method if you are wanting to run Chiron on in GPU mode (`pip install` version is coming). -``` -git clone https://github.com/haotianteng/chiron.git -cd chiron -``` -You will also need to install dependencies. - -For CPU-version: -``` -pip install tensorflow==1.0.1 -pip install h5py -``` -For GPU-version(Nvidia GPU required): -``` -pip install tensorflow-gpu==1.0.1 -pip install h5py -``` - -For alternate/detailed installation instructions for TensorFlow, see their [fantastic documentation](https://www.tensorflow.org/). - -## Basecall -### If installed from `pip`: -An example call to Chiron to run basecalling is: -``` -chiron call -i -o - -``` - -### If installed from GitHub: - -All Chiron functionality can be run from **entry.py** in the Chiron folder. (You might like to also add the path to Chiron into your path for ease of running). - -``` -python chiron/entry.py call -i -o - -``` - -### Test run - -We provide 5 sample fast5 files (courtesy of [nanonet](https://github.com/nanoporetech/nanonet)) in the GitHub repository which you can run a test on. These are located in `chiron/example_data/`. From inside the Chiron repository: -``` -python chiron/entry.py call -i chiron/example_folder/ -o -``` - - -### Output -`chiron call` will create five folders in `` called `raw`, `result`, `segments`, `meta`, and `reference`. - -* `result`: fastq/fasta files with the same name as the fast5 file they contain the basecalling result for. To create a single, merged version of these fasta files, try something like `paste --delimiter=\\n --serial result/*.fasta > merged.fasta` -* `raw`: Contains a file for each fast5 file with it's raw signal. This file format is an list of integers. i.e `544 554 556 571 563 472 467 487 482 513 517 521 495 504 500 520 492 506 ... ` -* `segments`: Contains the segments basecalled from each fast5 file. -* `meta`: Contains the meta information for each read (read length, basecalling rate etc.). Each file has the same name as it's fast5 file. -* `reference`: Contains the reference sequence (if any). - -### Output format -With -e flag to output fastq file(default) with quality score or fasta file. -Example: -chiron call -i -o -e fastq - - -chiron call -i -o -e fasta - -## Training -The default DNA model trained on R9.4 protocol with a mix of Lambda and E.coli dataset, if the basecalling result is not satisfying, you can train a model on your own training data set. - -#### Hardware request: -Recommend training on GPU with TensorFlow - usually 8GB RAM (GPU) is required. - -#### Prepare the training data set. -Using raw.py script to extract the signal and label from the re-squiggled fast5 file. -(For how to re-squiggle fast5 file, check [here, nanoraw re-squiggle](https://nanoraw.readthedocs.io/en/latest/resquiggle.html#example-commands)) - -#### If installed from `pip`: -``` -chiron export -i -o -``` - -or directly use the raw.py script in utils. - -``` -python chiron/utils/raw.py --input --output -``` -`.signal` file and correspond `.label` file, a typical file format: - -`.signal` file format: -`544 554 556 571 563 472 467 487 482 513 517 521 495 504 500 520 492 506 ...` -i.e the file must contain only one row/column of raw signal numbers. - -`.label` file format: -``` -70 174 A -174 184 T -184 192 A -192 195 G -195 204 C -204 209 A -209 224 C -... -``` - -Each line represents a DNA base pair in the Pore. -* 1st column: Start position of the current nucleotide, position related to the signal vector (index count starts from zero). -* 2nd column: End position of the current nucleotide. -* 3rd column: Nucleotide, for DNA: A, G, C, or T. Although, there is no reason you could not use other labels. - -#### Adjust Chiron parameters -Go in to `chiron/chiron_rcnn_train.py` and change the hyper parameters in the `FLAGS` class. - -```py -class Flags(): - def __init__(self): - self.home_dir = "/home/haotianteng/UQ/deepBNS/" - self.data_dir = self.home_dir + 'data/Lambda_R9.4/raw/' - self.log_dir = self.home_dir+'/chiron/log/' - self.sequence_len = 200 - self.batch_size = 100 - self.step_rate = 1e-3 - self.max_steps = 2500 - self.k_mer = 1 - self.model_name = 'crnn5+5_res_moving_norm' - self.retrain = False -``` - -`data_dir`: The folder containing your signal and label files. -`log_dir`: The folder where you want to save the model. -`sequence_len`: The length of the segment you want to separate the sequence into. Longer length requires larger RAM. -`batch_size`: The batch size. -`step_rate`: Learning rate of the optimizer. -`max_step`: Maximum step of the optimizer. -`k_mer`: Chiron supports learning based on k-mer instead of a single nucleotide, this should be an odd number, even numbers will cause an error. -`model_name`: The name of the model. The record will be stored in the directory `log_dir/model_name/` -`retrain`: If this is a new model, or you want to load the model you trained before. The model will be loaded from `log_dir/model_name/` - -### Train - -``` -source activate tensorflow -``` -#### If installed from `pip`: -``` -chiron train --data_dir --log_dir --model_name -``` - -or run directly by - -``` -python chiron/chiron_rcnn_train.py -``` +# Deepore +This modified version of Chiron allows us to visualise both the weights and the gradients to address training problem in Chiron, like local minima and bottlenecks during training that we have observed. diff --git a/Chiron+weight_visual/chiron/chiron_rcnn_train.py b/Chiron+weight_visual/chiron/chiron_rcnn_train.py index fe743f1..341fb99 100644 --- a/Chiron+weight_visual/chiron/chiron_rcnn_train.py +++ b/Chiron+weight_visual/chiron/chiron_rcnn_train.py @@ -13,6 +13,7 @@ #from rnn import rnn_layers from rnn import rnn_layers_one_direction import time,os +from summary import variable_summaries def save_model(): copy_tree(os.path.dirname(os.path.abspath(__file__)),FLAGS.log_dir+FLAGS.model_name+'/model') @@ -32,10 +33,13 @@ def loss(logits,seq_len,label): return loss def train_step(loss,global_step = None): - opt = tf.train.AdamOptimizer(FLAGS.step_rate).minimize(loss,global_step=global_step) + opt = tf.train.AdamOptimizer(FLAGS.step_rate) # opt = tf.train.GradientDescentOptimizer(FLAGS.step_rate).minimize(loss) # opt = tf.train.RMSPropOptimizer(FLAGS.step_rate).minimize(loss) # opt = tf.train.MomentumOptimizer(FLAGS.step_rate,0.9).minimize(loss) + grad = opt.compute_gradients(loss) + tf.summary.scalar('grad',tf.reduce_mean(grad[0][0])) + opt = opt.minimize(loss,global_step=global_step) return opt def prediction(logits,seq_length,label,top_paths=1): """ @@ -119,13 +123,14 @@ def run(args): if __name__ == "__main__": class Flags(): def __init__(self): - self.data_dir = '/media/haotianteng/Linux_ex/Nanopore_data/Lambda_R9.4/raw' - self.cache_dir = '/media/haotianteng/Linux_ex/Nanopore_data/Lambda_R9.4/cache' - self.log_dir = '/media/haotianteng/Linux_ex/GVM_model' + self.data_dir = '/home/docker/raw' #human + #self.data_dir = '/home/docker/ecoli' #ecoli + self.cache_dir = '/home/docker/out/cache' + self.log_dir = '/home/docker/out/logs' self.sequence_len = 300 - self.batch_size = 750 + self.batch_size = 64 self.step_rate = 1e-3 - self.max_steps = 20000 + self.max_steps = 10000 self.k_mer = 1 self.model_name = 'test' self.retrain =False diff --git a/Chiron+weight_visual/chiron/rnn.py b/Chiron+weight_visual/chiron/rnn.py index 4230408..997529b 100644 --- a/Chiron+weight_visual/chiron/rnn.py +++ b/Chiron+weight_visual/chiron/rnn.py @@ -60,5 +60,5 @@ def rnn_layers_one_direction(x,seq_length,training,hidden_num=200,layer_num = 3, lasth_rs = tf.reshape(lasth,[batch_size*max_time,hidden_num],name = 'lasth_rs') logits = tf.reshape(tf.nn.bias_add(tf.matmul(lasth_rs,weight_class),bias_class),[batch_size,max_time,class_n],name = "rnn_logits_rs") variable_summaries(weight_class) - variable_summaries(biases_out) + variable_summaries(bias_class) return logits diff --git a/Chiron+weight_visual/chiron/summary.py b/Chiron+weight_visual/chiron/summary.py index 56213fa..bc418d6 100644 --- a/Chiron+weight_visual/chiron/summary.py +++ b/Chiron+weight_visual/chiron/summary.py @@ -1,3 +1,5 @@ +import tensorflow as tf + def variable_summaries(var): """Attach a lot of summaries to a Tensor (for TensorBoard visualization).""" with tf.name_scope('summaries'): diff --git a/README.md b/README.md index c9a778f..d0716ec 100644 --- a/README.md +++ b/README.md @@ -29,17 +29,22 @@ we modified the docker from `https://github.com/anurag/fastai-course-1.git` ``` -nvidia-docker run -it \ +DATADIR=/data/nanopore + +nvidia-docker run \ + --rm -it \ --entrypoint /bin/zsh \ - -v /data/nanopore/new/fast5Dir/:/data \ - -p 8889:8888 \ + -v $DATADIR:/data \ + -p 8890:8888 \ + --name haruhi \ + -w /home/docker \ etheleon/chiron ``` To train deepore we need to run chiron_rcnn_train.py ``` -cd $HOME +export CUDA_VISIBLE_DEVICES="1" python Chiron/chiron/chiron_rcnn_train.py ```