'fieldset', '#title' => t('Enter Query Sequence'), '#description' => t('Enter one or more queries in the top text box or use the browse button to upload a file from your local disk. The file may contain a single sequence or a list of sequences. In both cases, the data must be in FASTA format. More information.. '), '#collapsible' => TRUE, '#collapsed' => FALSE, ); // Textfield for submitting a mult-FASTA query $form['query']['FASTA'] = array( '#type' => 'textarea', '#title' => t('Enter FASTA sequence(s)'), '#description'=>t('Enter query sequence(s) in the text area.'), ); // Upload a file as an alternative to enter a query sequence $form['#attributes']['enctype'] = 'multipart/form-data'; $form['query']['UPLOAD'] = array( '#prefix' => 'Or upload your query files: ', '#type' => 'file', '#description' => t('The file should be a plain-text FASTA file and not a .doc, .docx, etc. It cannot be greater than 10 Mb in size.'), ); // BLAST DATABASE //......................... $form['DB'] = array( '#type' => 'fieldset', '#title' => t('Choose Search Set'), '#description' => t('Choose from one of the nucleotide BLAST databases listed below. You can also use the browse button to upload a file from your local disk. The file may contain a single sequence or a list of sequences. '), '#collapsible' => TRUE, '#collapsed' => FALSE, ); $options = _DB_options(); $form['DB']['SELECT_DB'] = array( '#type' => 'select', '#title' => t('Nucleotide BLAST Databases:'), '#options' => $options, '#default_value' => 0, ); // Upload a file as an alternative to enter a query sequence $form['#attributes']['enctype'] = 'multipart/form-data'; $form['DB']['DBUPLOAD'] = array( '#prefix' => 'Or upload your own dataset: ', '#type' => 'file', '#description' => t('The file should be a plain-text FASTA file and not a .doc, .docx, etc. It cannot be greater than 10 Mb in size.'), ); // ALGORITHM PARAMETERS //......................... $form['ALG'] = array( '#type' => 'fieldset', '#title' => t('Algorithm parameters'), '#collapsible' => TRUE, '#collapsed' => TRUE, ); // General parameters $form['ALG']['GParam'] = array( '#type' => 'fieldset', '#title' => t('General parameters'), '#collapsible' => FALSE, ); $form['ALG']['GParam']['maxTarget'] = array( '#type' => 'select', '#title' => t('Max target sequences:'), '#options' => array( 0 => t('10'), 1 => t('50'), 2 => t('100'), 3 => t('250'), 4 => t('500'), 5 => t('1000'), 6 => t('5000'), 7 => t('10000'), 8 => t('20000'), ), '#default_value' => 2, '#description' => t('Select the maximum number of aligned sequences to display'), ); $form['ALG']['GParam']['shortQueries'] = array( '#type' => 'checkbox', '#title' => t('Automatically adjust parameters for short input sequences'), '#default_value' => TRUE, ); $form['ALG']['GParam']['eVal'] = array( '#type' => 'textfield', '#title' => t('e-Value (Expected Threshold)'), '#default_value' => 10, '#size' => 12, '#maxlength' => 20, '#description' => t('Expected number of chance matches in a random model. This number should be give in a decimal format. More Information | Expect value vedio tutorial'), ); $form['ALG']['GParam']['wordSize'] = array( '#type' => 'select', '#title' => t('Word size:'), '#options' => array( 0 => t('16'), 1 => t('20'), 2 => t('24'), 3 => t('28'), 4 => t('32'), 5 => t('48'), 6 => t('64'), 7 => t('128'), 8 => t('256'), ), '#default_value' => 3, '#description' => t('The length of the seed that initiates an alignment'), ); $form['ALG']['GParam']['qRange'] = array( '#type' => 'textfield', '#title' => t('Max matches in a query range'), '#default_value' => 0, '#size' => 12, '#maxlength' => 20, '#description' => t('Limit the number of matches to a query range. This option is useful if many strong matches to one part of a query may prevent BLAST from presenting weaker matches to another part of the query.'), ); // Scoring parameters $form['ALG']['SParam'] = array( '#type' => 'fieldset', '#title' => t('Scoring parameters'), '#collapsible' => FALSE, ); $form['ALG']['SParam']['M&MScores'] = array( '#type' => 'select', '#title' => t('Match/Mismatch Scores:'), '#options' => array( 0 => t('1,-2'), 1 => t('1,-3'), 2 => t('1,-4'), 3 => t('2,-3'), 4 => t('4,-5'), 5 => t('1,-1'), ), '#default_value' => 0, '#description' => t('Reward and penalty for matching and mismatching bases.'), ); $form['ALG']['SParam']['gapCost'] = array( '#type' => 'select', '#title' => t('Gap Costs:'), '#options' => array( 0 => t('Existence: 5 Extension: 2'), 1 => t('Existence: 2 Extension: 2'), 2 => t('Existence: 1 Extension: 2'), 3 => t('Existence: 0 Extension: 2'), 4 => t('Existence: 3 Extension: 1'), 5 => t('Existence: 2 Extension: 1'), 6 => t('Existence: 1 Extension: 1'), ), '#default_value' => 0, '#description' => t('Cost to create and extend a gap in an alignment. Linear costs are available only with megablast and are determined by the match/mismatch scores.'), ); // Submit $form['submit'] = array( '#type' => 'submit', '#default_value' => ' BLAST ', ); return $form; } /** * Form validation handler for blast_nucleotide_form(). * * @see blast_nucleotide_form(). */ function blast_nucleotide_form_validate($form, &$form_state) { // Validate FASTA seq in textfield $fastaSeq = $form_state['input']['FASTA']; if (isset($fastaSeq)) { if (_validateFasta($fastaSeq)){ form_set_error('nBLAST', t('Error: Failed to read the Blast query: Wrong format provided for FASTA nucleotide sequence')); } else { $form_state['qFlag'] = 'seqQuery'; } } // Validate query upload $upQuery = file_save_upload('UPLOAD', array('file_validate_extensions' => array('txt fasta fa fna')), FILE_EXISTS_RENAME); if ($upQuery) { $upQuery_uri = $upQuery->uri; $form_state['upQuery_path'] = drupal_realpath($upQuery_uri); $upQuery_content = file_get_contents($form_state['upQuery_path']); if (_validateFasta($upQuery_content)){ form_set_error('nBLAST', t('Error: Failed to upload the Blast query: Wrong format provided for FASTA nucleotide sequence')); } else { $form_state['qFlag'] = 'upQuery'; } } // Validate blast database upload $upDB = file_save_upload('DBUPLOAD', array('file_validate_extensions' => array('txt fasta fa fna')), FILE_EXISTS_RENAME); if ($upDB) { $upDB_uri = $upDB->uri; $form_state['upDB_path'] = drupal_realpath($upDB_uri); $upDB_content = file_get_contents($form_state['upDB_path']); if (_validateFasta($upDB_content)){ form_set_error('DB', t('Error: Failed to upload the Blast subject sequence file: Wrong format provided for FASTA nucleotide sequence')); } else { $form_state['dbFlag'] = 'upQuery'; } } else { $form_state['dbFlag'] = 'blastdb'; } } /** * Form submission handler for blast_nucleotide_form(). * * @see blast_nucleotide_form(). */ function blast_nucleotide_form_submit($form, &$form_state) { // SHOULDN'T BE HARDCODED!! exec("export BLASTDB=/home/Sequences/blast_dbs/"); $eVal = $form_state['values']['eVal']; $trgtKey = $form_state['values']['maxTarget']; $numAlign = $form['ALG']['GParam']['maxTarget']['#options'][$trgtKey]; $wsKey = $form_state['values']['wordSize']; $wordSize = $form['ALG']['GParam']['wordSize']['#options'][$wsKey]; // Expand Gap Cost key into open and extend penalties $gapKey = $form_state['values']['gapCost']; switch ($gapKey) { case 0: $gapOpen = 5; $gapExtend = 2; break; case 1: $gapOpen = 2; $gapExtend = 2; break; case 2: $gapOpen = 1; $gapExtend = 2; break; case 3: $gapOpen = 0; $gapExtend = 2; break; case 4: $gapOpen = 3; $gapExtend = 1; break; case 5: $gapOpen = 2; $gapExtend = 1; break; case 6: $gapOpen = 1; $gapExtend = 1; break; } // Epand Match/Mismatch option into penalty/reward values // @todo Amir: Is the switch supposed to be for $scoreKey? $scoreKey = $form_state['values']['M&MScores']; switch ($gapKey) { case 0: $penalty = -2; $reward = 1; break; case 1: $penalty = -3; $reward = 1; break; case 2: $penalty = -4; $reward = 1; break; case 3: $penalty = -3; $reward = 2; break; case 4: $penalty = -5; $reward = 4; break; case 5: $penalty = -1; $reward = 1; break; } // If the query was submitted via the texrfield then create a file containing it if ( isset($form_state['qFlag']) ) { if ( $form_state['qFlag'] == 'seqQuery' ) { $seq_content = $form_state['values']['FASTA']; $query = "/tmp/user__query_file.fasta"; file_put_contents ( $query , $seq_content); } elseif ( $form_state['qFlag'] == 'upQuery' ) { $query = $form_state['upQuery_path']; } } // If the BLAST database was uploaded then use it to run the BLAST if ( $form_state['dbFlag'] == 'upQuery') { $subjectSeq = $form_state['upDB_path']; $subSeqOut = drupal_basename($form_state['upDB_path']) . rand(0, 10000); $blast_subj_cmd = "blastn -query $query -subject $subjectSeq -out sites/default/files/$subSeqOut.blastn.html -evalue $eVal -word_size $wordSize -gapopen $gapOpen -gapextend $gapExtend -penalty $penalty -reward $reward -num_alignments 100 -html"; system($blast_subj_cmd); } // Otherwise, we are using one of the website provided BLAST databases so form the // BLAST command accordingly elseif ($form_state['dbFlag'] == 'blastdb') { $selected_db = $form_state['values']['SELECT_DB']; $blastdb_node = node_load($selected_db); $blastdb_path = $blastdb_node->db_path; $blastdb_human_name = $form['DB']['SELECT_DB']['#options'][$selected_db]; $subSeqOut = str_replace(' ','_', $blastdb_human_name) . rand(0, 10000); $blast_db_cmd = "blastn -task blastn -query $query -db $blastdb_path -out sites/default/files/$subSeqOut.blastn.xml -evalue $eVal -word_size $wordSize -gapopen $gapOpen -gapextend $gapExtend -penalty $penalty -reward $reward -num_alignments 100 -outfmt=5"; system($blast_db_cmd,$input); } // Redirect to the BLAST results page $path = "$subSeqOut.blastn.xml"; drupal_goto("blast/report/$path"); } /** * FASTA validating parser * * @param $sequence * A string of characters to be validated. A sequence in FASTA format begins with a single-line description, followed by lines of sequence data. * The description line is distinguished from the sequence data by a greater-than (">") symbol in the first column. * The word following the ">" symbol is the identifier of the sequence, and the rest of the line is the description (both are optional). * There should be no space between the ">" and the first letter of the identifier. The sequence ends if another line starting with a ">" appears; * this indicates the start of another sequence. * * @return * Return a boolean. 1 if the sequence does not pass the format valifation stage and 0 otherwise. * */ function _validateFasta($sequence) { $fastaIdRegEx = '/^>.*(\\n|\\r)/'; $fastaSeqRegEx = '/[^acgntuACGNTU\n\r]/'; if ( preg_match($fastaSeqRegEx,$sequence) && !(preg_match($fastaIdRegEx,$sequence)) ) { $flag = 1; } else { $flag = 0; } return $flag; } /** * Generate an array of BLAST database options based on existing nodes of type BlastDB * * @return * Return human readble names of the pre-existing blast databases */ function _DB_options() { $type = 'blastdb'; $nodes = node_load_multiple(array(), array('type'=> $type)); $options = array(); foreach ($nodes as $node) { if ( isset($node) && isset($node->db_dbtype) ) { if ( ($node->db_dbtype=='n') ) { $options[$node->nid] = $node->db_name; } } } asort($options); $options[0] = 'Select a Dataset'; return $options; }