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Update README.md
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mansikath committed Nov 26, 2023
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Expand Up @@ -20,20 +20,23 @@ fastqc SRR26624132_1.fastq SRR26624132_2.fastq
```
### Step 3: Trimming
Use Trimmomatic to trim low-quality bases:
- [Trimmomatic](http://www.usadellab.org/cms/?page=trimmomatic)

```bash
java -jar trimmomatic-0.39.jar PE -phred33 SRR26624132_1.fastq SRR26624132_2.fastq trim1_paired.fastq trim1_unpaired.fastq trim2_paired.fastq trim2_unpaired.fastq ILLUMINACLIP:TruSeq3-PE.fa:2:30:10 LEADING:3 TRAILING:3 SLIDINGWINDOW:4:15 MINLEN:36
```
### Step 4: Genome Assembly
Use Velvet for de novo genome assembly:

- [Velvet](http://www.ebi.ac.uk/~zerbino/velvet/)
or
Install Velvet (For macOS): brew install velvet
Install Velvet (For Ubuntu): sudo apt-get install velvet
```bash
velvetg ./velvet_output -clean yes -exp_cov 21 -cov_cutoff 2.81 -min_contig_lgth 200
```
### Step 5: BLAST Analysis
Use BLAST to compare assembled contigs with a reference genome:

- [blast executables](https://ftp.ncbi.nlm.nih.gov/blast/executables/blast+/LATEST/)
Install BLAST: brew install blast or sudo apt-get install ncbi-blast+
```bash
blastp -query ./velvet_output/contigs.fa -db ref_genome -out blast_result.txt -evalue 1e-04 -outfmt 6 -max_target_seqs 5 -num_threads 8
Expand All @@ -44,4 +47,4 @@ Retrieve top gene IDs from BLAST results:
```bash
cut -f 2 blast_result.txt | sort | uniq | head -n 10 > top_gene_ids.txt
```
Annotate genes using UniProt and PANTHER databases. Visit the PANTHER website and upload the gene list for functional annotation.
Annotate genes using UniProt and PANTHER databases. Visit the [PANTHER](https://www.pantherdb.org/) website and upload the gene list for functional annotation.

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