---
title: "BLAST Tutorial in R Markdown"
author: "Yaamini Venkataraman"
date: "10/11/2018"
output: html_document
---

In this tutorial, I'll use `blastx` in R Markdown. I will follow the same steps as the [BLAST Tutorial in Jupyter](https://github.com/fish546-2018/yaamini-virginica/tree/master/tutorials/2018-10-09-BLAST-Tutorial).

```{r setup, include=FALSE}
knitr::opts_chunk$set(echo = TRUE)
```

```{r}
sessionInfo()
```

```{bash}
pwd #Check present working directory
```

# Step 1: Create a BLAST Database

```{bash}
curl \
ftp://ftp.uniprot.org/pub/databases/uniprot/current_release/knowledgebase/complete/uniprot_sprot.fasta.gz \
> uniprot_sprot.fasta.gz #Download directory from Unprot Swiss-Prot
```

```{bash}
gunzip -k uniprot_sprot.fasta #Unzip the database, but keep the original file
```

```{bash}
ls -F
```

```{bash}
#Path to blast; path to input for database; specify database type; output name
/Applications/blast/ncbi-blast-2.2.18+/bin/makeblastdb \
-in uniprot_sprot.fasta \
-dbtype prot \
-out uniprot_sprot_20181011
```
 
```{bash}
ls -F #Confirm database creation worked
```

# Step 2: Obtain the query sequence

```{bash}
curl http://eagle.fish.washington.edu/cnidarian/Ab_4denovo_CLC6_a.fa \
> Ab_4denovo_CLC6_a.fa
```

```{bash}
head Ab_4denovo_CLC6_a.fa #Confirm download
```

```{bash}
grep -c ">" Ab_4denovo_CLC6_a.fa #Count the number of sequences
```

# Step 3: Run `blastx`

To run blastx I need the following:

1. Path to blastx
2. Path to query
3. Path to database
4. Output file name
5. Maximum e-value to use for hits
6. Number of CPUs to use
7. Only produce 1 hit per query (may not be the best hit, but it is the first hit the algorithm finds...this is different than the web interface that finds the best hit)
8. Output format 6 specifies tabular format

```{bash}
/Applications/blast/ncbi-blast-2.2.18+/bin/makeblastdb \
-query Ab_4denovo_CLC6_a.fa \
-db uniprot_sprot_20181010  \
-out Ab_4-uniprot_blastx.tab \
-evalue 1E-20 \
-num_threads 4 \
-max_target_seqs 1 \
-outfmt 6
```

```{bash}
head Ab_4-uniprot_blastx.tab #Confirm output was created
```

```{bash}
wc -l Ab_4-uniprot_blastx.tab #Count the number of hits
```