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Showing sgRNA sequences on hover in CRISPRessoPro (pinellolab#432)
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* Passing sgRNA sequences to regular and Batch D3 plots (#73)

* Sam/try plots (#71)

* Fix batch mode pandas warning. (#70)

* refactor to call method on DataFrame, rather than Series.
Removes warning.

* Fix pandas future warning in CRISPRessoWGS

---------

Co-authored-by: Cole Lyman <cole@colelyman.com>

* Functional

* Cole/fix status file name (#69)

* Update config file logging messages

This removes printing the exception (which is essentially a duplicate),
and adds a condition if no config file was provided. Also changes `json`
to `config` so that it is more clear.

* Fix divide by zero when no amplicons are present in Batch mode

* Don't append file_prefix to status file name

* Place status files in output directories

* Update tests branch for file_prefix addition

* Load D3 and plotly figures with pro with multiple amplicons

* Update batch

* Fix bug in CRISPRessoCompare with pointing to report datas with file_prefix

Before this fix, when using a file_prefix the second run that was compared
would not be displayed as a data in the first figure of the report.

* Import CRISPRessoPro instead of importing the version

When installed via conda, the version is not available

* Remove `get_amplicon_output` unused function from CRISPRessoCompare

Also remove unused argparse import

* Implement `get_matching_allele_files` in CRISPRessoCompare and accompanying unit tests

* Allow for matching of multiple guides in the same amplicon

* Fix pandas FutureWarning

* Change test branch back to master

---------

Co-authored-by: Sam <snic9004@gmail.com>

* Try catch all futures

* Fix test fail plots

* Point test to try-plots

* Fix d3 not showing and plotly mixing with matplotlib

* Use logger for warnings and debug statements

* Point tests back at master

---------

Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

* Sam/fix plots (#72)

* Fix batch mode pandas warning. (#70)

* refactor to call method on DataFrame, rather than Series.
Removes warning.

* Fix pandas future warning in CRISPRessoWGS

---------

Co-authored-by: Cole Lyman <cole@colelyman.com>

* Functional

* Cole/fix status file name (#69)

* Update config file logging messages

This removes printing the exception (which is essentially a duplicate),
and adds a condition if no config file was provided. Also changes `json`
to `config` so that it is more clear.

* Fix divide by zero when no amplicons are present in Batch mode

* Don't append file_prefix to status file name

* Place status files in output directories

* Update tests branch for file_prefix addition

* Load D3 and plotly figures with pro with multiple amplicons

* Update batch

* Fix bug in CRISPRessoCompare with pointing to report datas with file_prefix

Before this fix, when using a file_prefix the second run that was compared
would not be displayed as a data in the first figure of the report.

* Import CRISPRessoPro instead of importing the version

When installed via conda, the version is not available

* Remove `get_amplicon_output` unused function from CRISPRessoCompare

Also remove unused argparse import

* Implement `get_matching_allele_files` in CRISPRessoCompare and accompanying unit tests

* Allow for matching of multiple guides in the same amplicon

* Fix pandas FutureWarning

* Change test branch back to master

---------

Co-authored-by: Sam <snic9004@gmail.com>

* Try catch all futures

* Fix test fail plots

* Fix d3 not showing and plotly mixing with matplotlib

---------

Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

* Remove token from integration tests file

* Provide sgRNA_sequences to plot_nucleotide_quilt plots

* Passing sgRNA_sequences to plot

* Refactor check for determining when to use CRISPREssoPro or matplotlib for Batch plots

* Add max-height to Batch report samples

* Change testing branch

* Fix wrong check for large Batch plots

* Update integration_tests.yml to point back at master

---------

Co-authored-by: Samuel Nichols <Snic9004@gmail.com>
Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

* Push new releases to ECR (#74)

* Create aws_ecr.yml (#1)

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* us-east-1

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Update aws_ecr.yml

* Fix d3 sgRNA sequences (#76)

* Pass correct sgRNA_sequences to d3 plot

* Pass correct sgRNA sequence to prime editor plot for d3

* Resize plotly (#75)

* Sam/try plots (#71)

* Fix batch mode pandas warning. (#70)

* refactor to call method on DataFrame, rather than Series.
Removes warning.

* Fix pandas future warning in CRISPRessoWGS

---------

Co-authored-by: Cole Lyman <cole@colelyman.com>

* Functional

* Cole/fix status file name (#69)

* Update config file logging messages

This removes printing the exception (which is essentially a duplicate),
and adds a condition if no config file was provided. Also changes `json`
to `config` so that it is more clear.

* Fix divide by zero when no amplicons are present in Batch mode

* Don't append file_prefix to status file name

* Place status files in output directories

* Update tests branch for file_prefix addition

* Load D3 and plotly figures with pro with multiple amplicons

* Update batch

* Fix bug in CRISPRessoCompare with pointing to report datas with file_prefix

Before this fix, when using a file_prefix the second run that was compared
would not be displayed as a data in the first figure of the report.

* Import CRISPRessoPro instead of importing the version

When installed via conda, the version is not available

* Remove `get_amplicon_output` unused function from CRISPRessoCompare

Also remove unused argparse import

* Implement `get_matching_allele_files` in CRISPRessoCompare and accompanying unit tests

* Allow for matching of multiple guides in the same amplicon

* Fix pandas FutureWarning

* Change test branch back to master

---------

Co-authored-by: Sam <snic9004@gmail.com>

* Try catch all futures

* Fix test fail plots

* Point test to try-plots

* Fix d3 not showing and plotly mixing with matplotlib

* Use logger for warnings and debug statements

* Point tests back at master

---------

Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

* Sam/fix plots (#72)

* Fix batch mode pandas warning. (#70)

* refactor to call method on DataFrame, rather than Series.
Removes warning.

* Fix pandas future warning in CRISPRessoWGS

---------

Co-authored-by: Cole Lyman <cole@colelyman.com>

* Functional

* Cole/fix status file name (#69)

* Update config file logging messages

This removes printing the exception (which is essentially a duplicate),
and adds a condition if no config file was provided. Also changes `json`
to `config` so that it is more clear.

* Fix divide by zero when no amplicons are present in Batch mode

* Don't append file_prefix to status file name

* Place status files in output directories

* Update tests branch for file_prefix addition

* Load D3 and plotly figures with pro with multiple amplicons

* Update batch

* Fix bug in CRISPRessoCompare with pointing to report datas with file_prefix

Before this fix, when using a file_prefix the second run that was compared
would not be displayed as a data in the first figure of the report.

* Import CRISPRessoPro instead of importing the version

When installed via conda, the version is not available

* Remove `get_amplicon_output` unused function from CRISPRessoCompare

Also remove unused argparse import

* Implement `get_matching_allele_files` in CRISPRessoCompare and accompanying unit tests

* Allow for matching of multiple guides in the same amplicon

* Fix pandas FutureWarning

* Change test branch back to master

---------

Co-authored-by: Sam <snic9004@gmail.com>

* Try catch all futures

* Fix test fail plots

* Fix d3 not showing and plotly mixing with matplotlib

---------

Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

* Remove token from integration tests file

* Pass div id for plotly

* Remove debug

---------

Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
Co-authored-by: Cole Lyman <cole@colelyman.com>

---------

Co-authored-by: Trevor Martin <60452953+trevormartinj7@users.noreply.github.com>
Co-authored-by: Samuel Nichols <Snic9004@gmail.com>
Co-authored-by: mbowcut2 <55161542+mbowcut2@users.noreply.github.com>
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47 changes: 47 additions & 0 deletions .github/workflows/aws_ecr.yml
Original file line number Diff line number Diff line change
@@ -0,0 +1,47 @@
name: Push Docker image to Amazon ECR

on:
release:
types:
- edited
- released
branches:
- master

jobs:
build-and-push:
name: Build and Push Docker image
runs-on: ubuntu-latest

steps:
- name: Checkout
uses: actions/checkout@v2

- id: get_version
name: Get version
uses: jannemattila/get-version-from-tag@v3

- name: Configure AWS credentials
uses: aws-actions/configure-aws-credentials@v1
with:
aws-access-key-id: ${{ secrets.AWS_ACCESS_KEY_ID }}
aws-secret-access-key: ${{ secrets.AWS_SECRET_ACCESS_KEY }}
aws-region: us-east-1

- name: Login to Amazon ECR
id: login-ecr
uses: aws-actions/amazon-ecr-login@v1

- name: Build, tag, and push the image to Amazon ECR
id: build-image
env:
AWS_ACCOUNT: ${{ secrets.AWS_ACCOUNT_ID }}
ECR_REPOSITORY: 'crispresso2'
AWS_REGION: 'us-east-1'
IMAGE_TAG: ${{ steps.get_version.outputs.version }}
run: |
# Build a docker container and push it to ECR
docker build -t $AWS_ACCOUNT.dkr.ecr.$AWS_REGION.amazonaws.com/$ECR_REPOSITORY:$IMAGE_TAG .
echo "Pushing image to ECR..."
docker push $AWS_ACCOUNT.dkr.ecr.$AWS_REGION.amazonaws.com/$ECR_REPOSITORY:$IMAGE_TAG
echo "::set-output name=image::$AWS_ACCOUNT.dkr.ecr.$AWS_REGION.amazonaws.com/$ECR_REPOSITORY:$IMAGE_TAG"
8 changes: 8 additions & 0 deletions CRISPResso2/CRISPRessoAggregateCORE.py
Original file line number Diff line number Diff line change
Expand Up @@ -614,12 +614,14 @@ def main():
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_titles'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_labels'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_datas'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_divs'] = {}

crispresso2_info['results']['general_plots']['allele_modification_line_plot_names'] = []
crispresso2_info['results']['general_plots']['allele_modification_line_plot_paths'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_titles'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_labels'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_datas'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_divs'] = {}
if guides_all_same:
sgRNA_intervals = [consensus_sgRNA_intervals] * modification_frequency_summary_df.shape[0]
else:
Expand All @@ -645,11 +647,13 @@ def main():
plot_name = 'CRISPRessoAggregate_percentage_of_{0}_across_alleles_{1}_heatmap'.format(modification_type.lower(), amplicon_name)
plot_path = '{0}.html'.format(_jp(plot_name))

heatmap_div_id = '{0}-allele-modification-heatmap-{1}'.format(amplicon_name.lower(), modification_type.lower())
allele_modification_heatmap_input = {
'sample_values': modification_df,
'sample_sgRNA_intervals': sgRNA_intervals,
'plot_path': plot_path,
'title': modification_type,
'div_id': heatmap_div_id,
}
plot(
CRISPRessoPlot.plot_allele_modification_heatmap,
Expand All @@ -671,15 +675,18 @@ def main():
),
),
]
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_divs'][plot_name] = heatmap_div_id

plot_name = 'CRISPRessoAggregate_percentage_of_{0}_across_alleles_{1}_line'.format(modification_type.lower(), amplicon_name)
plot_path = '{0}.html'.format(_jp(plot_name))

line_div_id = '{0}-allele-modification-line-{1}'.format(amplicon_name.lower(), modification_type.lower())
allele_modification_line_input = {
'sample_values': modification_df,
'sample_sgRNA_intervals': sgRNA_intervals,
'plot_path': plot_path,
'title': modification_type,
'div_id': line_div_id,
}
plot(
CRISPRessoPlot.plot_allele_modification_line,
Expand All @@ -700,6 +707,7 @@ def main():
),
),
]
crispresso2_info['results']['general_plots']['allele_modification_line_plot_divs'][plot_name] = line_div_id

crispresso2_info['results']['general_plots']['window_nuc_pct_quilt_plot_names'] = window_nuc_pct_quilt_plot_names
crispresso2_info['results']['general_plots']['nuc_pct_quilt_plot_names'] = nuc_pct_quilt_plot_names
Expand Down
55 changes: 44 additions & 11 deletions CRISPResso2/CRISPRessoBatchCORE.py
Original file line number Diff line number Diff line change
Expand Up @@ -49,6 +49,33 @@ def check_library(library_name):
np = check_library('numpy')


def should_plot_large_plots(num_rows, c2pro_installed, use_matplotlib, large_plot_cutoff=300):
"""Determine if large plots should be plotted.
Parameters
----------
num_rows : int
Number of rows in the dataframe.
c2pro_installed : bool
Whether CRISPRessoPro is installed.
use_matplotlib : bool
Whether to use matplotlib when CRISPRessoPro is installed, i.e. value
of `--use_matplotlib`.
large_plot_cutoff : int, optional
Number of samples at which to not plot large plots with matplotlib.
Note that each sample has 6 rows in the datafame. Defaults to 300.
Returns
-------
bool
Whether to plot large plots.
"""
return (
(not use_matplotlib and c2pro_installed)
or (num_rows / 6) < large_plot_cutoff
)


def main():
try:
start_time = datetime.now()
Expand Down Expand Up @@ -388,14 +415,14 @@ def main():
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_titles'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_labels'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_datas'] = {}
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_divs'] = {}

crispresso2_info['results']['general_plots']['allele_modification_line_plot_names'] = []
crispresso2_info['results']['general_plots']['allele_modification_line_plot_paths'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_titles'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_labels'] = {}
crispresso2_info['results']['general_plots']['allele_modification_line_plot_datas'] = {}

large_plot_cutoff = 300
crispresso2_info['results']['general_plots']['allele_modification_line_plot_divs'] = {}

percent_complete_start, percent_complete_end = 90, 99
if all_amplicons:
Expand Down Expand Up @@ -580,7 +607,7 @@ def main():
sub_modification_percentage_summary_filename = _jp(amplicon_plot_name + 'Modification_percentage_summary_around_sgRNA_'+sgRNA+'.txt')
sub_modification_percentage_summary_df.to_csv(sub_modification_percentage_summary_filename, sep='\t', index=None)

if not args.suppress_plots and not args.suppress_batch_summary_plots and (nucleotide_percentage_summary_df.shape[0] / 6) < large_plot_cutoff:
if not args.suppress_plots and not args.suppress_batch_summary_plots and should_plot_large_plots(sub_nucleotide_percentage_summary_df.shape[0], C2PRO_INSTALLED, args.use_matplotlib):
# plot for each guide
# show all sgRNA's on the plot
sub_sgRNA_intervals = []
Expand Down Expand Up @@ -614,6 +641,7 @@ def main():
'fig_filename_root': f'{this_window_nuc_pct_quilt_plot_name}.json' if not args.use_matplotlib and C2PRO_INSTALLED else this_window_nuc_pct_quilt_plot_name,
'save_also_png': save_png,
'sgRNA_intervals': sub_sgRNA_intervals,
'sgRNA_sequences': consensus_guides,
'quantification_window_idxs': include_idxs,
'custom_colors': custom_config['colors'],
}
Expand All @@ -628,7 +656,7 @@ def main():
crispresso2_info['results']['general_plots']['summary_plot_labels'][plot_name] = 'Composition of each base around the guide ' + sgRNA + ' for the amplicon ' + amplicon_name
crispresso2_info['results']['general_plots']['summary_plot_datas'][plot_name] = [('Nucleotide frequencies', os.path.basename(nucleotide_frequency_summary_filename)), ('Modification frequencies', os.path.basename(modification_frequency_summary_filename))]

if args.base_editor_output and (sub_nucleotide_percentage_summary_df.shape[0] / 6) < large_plot_cutoff:
if args.base_editor_output and should_plot_large_plots(sub_nucleotide_percentage_summary_df.shape[0], False, args.use_matplotlib):
this_window_nuc_conv_plot_name = _jp(amplicon_plot_name + 'Nucleotide_conversion_map_around_sgRNA_'+sgRNA)
conversion_map_input = {
'nuc_pct_df': sub_nucleotide_percentage_summary_df,
Expand Down Expand Up @@ -656,14 +684,15 @@ def main():
]
# done with per-sgRNA plots

if not args.suppress_plots and not args.suppress_batch_summary_plots: # plot the whole region
if not args.suppress_plots and not args.suppress_batch_summary_plots and should_plot_large_plots(nucleotide_percentage_summary_df.shape[0], C2PRO_INSTALLED, args.use_matplotlib): # plot the whole region
this_nuc_pct_quilt_plot_name = _jp(amplicon_plot_name.replace('.', '') + 'Nucleotide_percentage_quilt')
nucleotide_quilt_input = {
'nuc_pct_df': nucleotide_percentage_summary_df,
'mod_pct_df': modification_percentage_summary_df,
'fig_filename_root': f'{this_nuc_pct_quilt_plot_name}.json' if not args.use_matplotlib and C2PRO_INSTALLED else this_nuc_pct_quilt_plot_name,
'save_also_png': save_png,
'sgRNA_intervals': consensus_sgRNA_intervals,
'sgRNA_sequences': consensus_guides,
'quantification_window_idxs': include_idxs,
'custom_colors': custom_config['colors'],
}
Expand All @@ -679,7 +708,7 @@ def main():
crispresso2_info['results']['general_plots']['summary_plot_titles'][plot_name] = ''
crispresso2_info['results']['general_plots']['summary_plot_labels'][plot_name] = 'Composition of each base for the amplicon ' + amplicon_name
crispresso2_info['results']['general_plots']['summary_plot_datas'][plot_name] = [('Nucleotide frequencies', os.path.basename(nucleotide_frequency_summary_filename)), ('Modification frequencies', os.path.basename(modification_frequency_summary_filename))]
if args.base_editor_output and (sub_nucleotide_percentage_summary_df.shape[0] / 6) < large_plot_cutoff:
if args.base_editor_output and should_plot_large_plots(nucleotide_percentage_summary_df.shape[0], False, args.use_matplotlib):
this_nuc_conv_plot_name = _jp(amplicon_plot_name + 'Nucleotide_conversion_map')
conversion_map_input = {
'nuc_pct_df': nucleotide_percentage_summary_df,
Expand All @@ -706,7 +735,7 @@ def main():
crispresso2_info['results']['general_plots']['summary_plot_datas'][plot_name] = [('Nucleotide frequencies', os.path.basename(nucleotide_frequency_summary_filename)), ('Modification frequencies', os.path.basename(modification_frequency_summary_filename))]

else: # guides are not the same
if not args.suppress_plots and not args.suppress_batch_summary_plots:
if not args.suppress_plots and not args.suppress_batch_summary_plots and should_plot_large_plots(nucleotide_percentage_summary_df.shape[0], C2PRO_INSTALLED, args.use_matplotlib):
this_nuc_pct_quilt_plot_name = _jp(amplicon_plot_name.replace('.', '') + 'Nucleotide_percentage_quilt')
nucleotide_quilt_input = {
'nuc_pct_df': nucleotide_percentage_summary_df,
Expand All @@ -724,7 +753,7 @@ def main():
nuc_pct_quilt_plot_names.append(plot_name)
crispresso2_info['results']['general_plots']['summary_plot_labels'][plot_name] = 'Composition of each base for the amplicon ' + amplicon_name
crispresso2_info['results']['general_plots']['summary_plot_datas'][plot_name] = [('Nucleotide frequencies', os.path.basename(nucleotide_frequency_summary_filename)), ('Modification frequencies', os.path.basename(modification_frequency_summary_filename))]
if args.base_editor_output and (sub_nucleotide_percentage_summary_df.shape[0] / 6) < large_plot_cutoff:
if args.base_editor_output and should_plot_large_plots(nucleotide_percentage_summary_df.shape[0], False, args.use_matplotlib):
this_nuc_conv_plot_name = _jp(amplicon_plot_name + 'Nucleotide_percentage_quilt')
conversion_map_input = {
'nuc_pct_df': nucleotide_percentage_summary_df,
Expand All @@ -745,7 +774,7 @@ def main():
crispresso2_info['results']['general_plots']['summary_plot_datas'][plot_name] = [('Nucleotide frequencies', os.path.basename(nucleotide_frequency_summary_filename)), ('Modification frequencies', os.path.basename(modification_frequency_summary_filename))]

# allele modification frequency heatmap and line plots
if C2PRO_INSTALLED and not args.use_matplotlib and not args.suppress_plots and not args.suppress_batch_summary_plots and (nucleotide_percentage_summary_df.shape[0] / 6) < large_plot_cutoff:
if C2PRO_INSTALLED and not args.use_matplotlib and not args.suppress_plots and not args.suppress_batch_summary_plots:
if guides_all_same:
sgRNA_intervals = [consensus_sgRNA_intervals] * modification_frequency_summary_df.shape[0]
else:
Expand All @@ -771,12 +800,13 @@ def main():
plot_name = 'CRISPRessoBatch_percentage_of_{0}_across_alleles_{1}_heatmap'.format(modification_type.lower(), amplicon_name)
plot_path = '{0}.html'.format(_jp(plot_name))

heatmap_div_id = '{0}-allele-modification-heatmap-{1}'.format(amplicon_name.lower(), modification_type.lower())
allele_modification_heatmap_input = {
'sample_values': modification_df,
'sample_sgRNA_intervals': sgRNA_intervals,
'plot_path': plot_path,
'title': modification_type,
'amplicon_name': amplicon_name,
'div_id': heatmap_div_id,
}
debug('Plotting allele modification heatmap for {0}'.format(amplicon_name))
plot(
Expand All @@ -799,16 +829,18 @@ def main():
),
),
]
crispresso2_info['results']['general_plots']['allele_modification_heatmap_plot_divs'][plot_name] = heatmap_div_id

plot_name = 'CRISPRessoBatch_percentage_of_{0}_across_alleles_{1}_line'.format(modification_type.lower(), amplicon_name)
plot_path = '{0}.html'.format(_jp(plot_name))

line_div_id = '{0}-allele-modification-line-{1}'.format(amplicon_name.lower(), modification_type.lower())
allele_modification_line_input = {
'sample_values': modification_df,
'sample_sgRNA_intervals': sgRNA_intervals,
'plot_path': plot_path,
'title': modification_type,
'amplicon_name': amplicon_name,
'div_id': line_div_id,
}
debug('Plotting allele modification line plot for {0}'.format(amplicon_name))
plot(
Expand All @@ -831,6 +863,7 @@ def main():
),
),
]
crispresso2_info['results']['general_plots']['allele_modification_line_plot_divs'][plot_name] = line_div_id
#end if amp_found_count > 0 (how many folders had information for this amplicon)
#end per-amplicon analysis

Expand Down
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