Lab page @ http://krishna.gs.washington.edu/index.html

What is RNA-Seq?

RNA-Seq is a next-generation sequencing (NGS) technology used to study the transcriptome—the complete set of RNA molecules in a cell. It quantifies gene expression, detects novel transcripts, and captures alternative splicing events with high sensitivity and resolution.

Workflow for RNA-Seq Analysis

RNA-Seq analysis involves several stages, each requiring computational tools and expertise.

1. Experimental Design and Data Acquisition

Before diving into analysis, bioinformaticians should consider:

• Biological Replicates: Ensure statistical power to detect meaningful differences.
• Sequencing Depth: Align sequencing depth to study objectives (e.g., higher depth for low-abundance transcripts).
• Paired-End vs. Single-End: Paired-end sequencing provides more detailed information on transcript structure.

Once sequencing is complete, raw data is provided in FASTQ format, containing sequence reads and quality scores.

2. Quality Control and Preprocessing

Quality control (QC) ensures data integrity. Tools such as FastQC evaluate metrics like base quality, GC content, and adapter contamination.

Preprocessing Steps:

• Trimming: Tools like Trimmomatic or Cutadapt remove low-quality bases and adapter sequences.
• Filtering: Discard reads below a certain quality threshold or length.

3. Read Alignment

Reads are mapped to a reference genome or transcriptome to determine their origin. Alignment tools include:

• HISAT2: Handles large genomes efficiently and supports spliced alignments.
• STAR: High-speed aligner optimized for RNA-Seq.
• Bowtie2: Suitable for short-read alignment.

Output: A SAM/BAM file containing aligned reads.

4. Transcript Assembly and Quantification

This step involves identifying transcripts and quantifying their expression levels. Tools used include:

• StringTie: Assembles and quantifies transcripts from aligned reads.
• Salmon/Kallisto: Perform pseudo-alignment for rapid and accurate quantification.

Expression levels are typically measured as TPM (transcripts per million) or FPKM (fragments per kilobase of transcript per million mapped reads).

5. Differential Expression Analysis

To identify genes with altered expression between conditions, bioinformaticians use tools such as:

• DESeq2: Accounts for data normalization and variability.
• edgeR: Handles overdispersed count data efficiently.
• Limma-voom: Combines linear modeling with RNA-Seq count data.

The output includes a list of differentially expressed genes (DEGs) with statistical significance and fold-change values.

6. Functional Annotation and Pathway Analysis

Understanding the biological significance of DEGs involves:

• Gene Ontology (GO) Analysis: Tools like DAVID or clusterProfiler categorize genes based on their biological functions.
• Pathway Enrichment Analysis: Identifies pathways enriched in DEGs using tools like KEGG, Reactome, or GSEA.

7. Visualization

Visualizing results enhances interpretability. Common visualizations include:

• Heatmaps: Show expression patterns across samples (e.g., pheatmap).
• Volcano Plots: Highlight significant DEGs (e.g., ggplot2).
• PCA/UMAP: Assess sample clustering and variability (e.g., Seurat).

Challenges in RNA-Seq Analysis

1. Batch Effects: Technical variability can confound biological signals. Combat this with normalization techniques or batch-correction tools like ComBat.
2. Low-Quality Samples: Poor-quality RNA impacts downstream analyses.
3. Computational Complexity: RNA-Seq generates massive datasets, requiring robust computing resources and optimized pipelines.

Key Tools and Resources

• Bioconductor: A treasure trove of R packages for RNA-Seq analysis.
• Galaxy: A web-based platform for running RNA-Seq workflows.
• Nextflow/Snakemake: Workflow management tools to streamline analyses.

Applications of RNA-Seq

RNA-Seq is used in diverse research areas, including:

• Cancer Transcriptomics: Identifying tumor-specific expression profiles.
• Developmental Biology: Studying dynamic transcriptome changes.
• Drug Discovery: Screening genes modulated by therapeutic compounds.

Conclusion

RNA-Seq analysis is a cornerstone of modern transcriptomics, offering bioinformaticians a versatile toolkit for unraveling gene expression and regulation. Mastering RNA-Seq workflows and tools empowers researchers to transform raw sequencing data into biological discoveries.

Whether you’re investigating disease mechanisms, exploring cellular pathways, or developing new therapeutics, RNA-Seq is a powerful ally in your bioinformatics arsenal.

(A) Regulatory System Analysis with respect to microRNAs

(B) Computational Epigenomics & Regulomics:

(C) Computational issues with Next Generation Sequencing:

Department of Biotechnology,
Institute of Himalyan Bioresources Technology
CSIR, Palampur(Himachal Pradesh), India.
Email: ravishihbt.res.in; ravish9gmail.com

More @ http://scbb.ihbt.res.in/SCBB_dept/Lab_Member.php

More @ http://www.bioinf.manchester.ac.uk/dbbrowser/index.php

1. Project Junior Research Fellow / Project Assistant

Last Date: 11th Nov 2013

Qualification B.Tech (Comp. Sci.), B.Tech/M.Tech (Bioinformatics), MCA, M.Sc. (Mathematics/Statistics)

Desirable Qualifications: Programming in FORTRAN/ C /PERL, Web application technologies

Upper Age limit 28

Rs.12000 / Rs.16000 (as sanctioned by the funding agency)

General terms and conditions:

Positions are purely temporary and co-terminus with the project.

HRDG (CSIR) prevailing guidelines are applicable these positions.

All categories of applicants are required to submit online application.

Enhancement of stipend to Project JRF to Project SRF will be with the due recommendation of Principal Investigator and approval of the Director on the evaluation of the 3 member Standing Committee consisting of Chairperson at the level of Chief Scientist, Coordinator of the JRFs/RAs/PDFs and the Principal Investigator of the Project.

The age relaxation as per HRDG (CSIR) norms: SC/ST/OBC/Women/Physically Handicapped persons – five years.

The Stipend normally be fixed at Rs.22000/- for Research Associates/Post Doc. Fellows. However, a selected RA/PDF may be placed in the higher start of stipend if there is ample justification and such recommendation is made by the Selection Committee. Based on the recommendation with justification by the PI and approval of the Director, person getting stipend at lower rate may be elevated to higher rate subject to availability of the funds in the project.

Recruitment will be based on initial screening based on qualifications and experience criteria and also based on suitability of the candidates to the nature of research project. This screening will be followed by written test followed / interview. After completing this process, candidates will be shortlisted and appointed in specific project subjects as and when appropriate positions become available. The pool of selected candidates will be valid for six months.

Remunerations indicate are maximum admissible and will depend upon the availability of funds and subject to conditions applicable to projects from different funding agencies at the time of recruitment.

Apply : http://www.ccmb.res.in/positions/projects/temp_positions.php

Form download : http://www.ccmb.res.in/positions/projects/oct-2013/pdf_download.php

Lab page: http://www.oeb.harvard.edu/faculty/edwards/index.html

Last Date for Submitting Application: 25th November 2013

1. Senior Scientist: (01 position)
Pay Scale: Rs.40, 000/-
Qualifications: PhD/ Post Doctoral with Experience in CADD

2. Junior Scientist (10 positions)
Pay Scale: Rs. 22,000/-
Qualifications: MPhil / Masters Degree in Bioinformatics / Computational Biology / CADD / Ayurveda

3. Technical Assistant (01+01 positions)
Pay Scale: Rs.12,000/-
Qualifications: 1. BSc Computer Science/ MCA
Qualifications: 2. MSc Biotechnology / MSc Microbiology

4. Programmer (01 position)
Pay Scale: Rs.20,000/-
Qualifications: MSc Computer Science/ MCA / B Tech (Experience in MATLAB, C, C++) Industrial experience is desirable

5. Teaching Assistant (03 positions)
Pay Scale: Rs.10,000/-
Qualifications: MSc in Bioinformatics

6. Administration Assistant (02 positions)
Pay Scale: Rs.8,000/-
Qualifications: Degree + PGDCA

The Selection process comprises of written test and interview. Positions are purely temporary (initially for the period of one year) and co-terminus with the project. For more details mail to: cbi.uok [at] gmail.com

More detail @ https://sites.google.com/site/centreforbioinformatics/announcements/applicationsinvitedforapplicationforai-caddproject

WALK-IN-INTERVIEW

The following vacancies shall be filled purely on adhoc basis under Non-Institutional adhoc project “Bioinformatics in ICMR Institutes” funded by Indian Council of Medical Research at Vector Control Research Centre, Puducherry, to be renewed annually and filled through Walk-in-Interview as indicated below. Candidates who wish to appear for the Walk-in-Interview can download the application format given in the website of Vector Control Research Centre (www.vcrc.res.in). Duly filled in application along with attested copies of certificate should be submitted at time of interview.

Date & Time : 05.12.2013 at 9.00 AM – Scientist-C (Non-Medical)

05.12.2013 at 1.30 PM – Scientist-B (Non-Medical)
06.12.2013 at 9.00 AM – Technical Assistant (Research Assistant)
06.12.2013 at 1.30 PM – Multi Tasking Staff (General)

Place : Vector Control Research Centre, Puducherry

Project entitled : Biomedical Informatics Centres of ICMR

1. Scientist - C (Non-Medical) Number of post – ONE

Essential qualification

B.E./ B. Tech. Degree in Bioinformatics/ Computational Biology from a recognized University with 6 years experience in the relevant field OR

First class Master’s Degree and Ph.D. Degree in Bioinformatics/ Computational Biology from a recognized University OR

First class Master’s Degree in Bioinformatics/ Computational Biology from a recognized University with 4 years R & D experience in the related subjects as mentioned above OR

Second class Master’s Degree + Ph.D. in Bioinformatics/ Computational Biology from a recognized University with 4 years research experience in bio-medical subjects

Age: Not exceeding 40 years Consolidated Salary – Rs.39,960/- p.m. + HRA as
admissible

Desirable qualification (i) Post-doctorate in Bioinformatics/ Computational Biology or M.E. / M. Tech. Degree in Bioinformatics/ Computational Biology from a recognized University for candidates with First Class relevant degree.

(ii) Additional post-doctoral research / teaching experience in Bioinformatics/Computational Biology in recognized Institute(s).

(iii) Knowledge of computer applications or data management

Job requirements i) To apply Bioinformatics / Computational Biology tools in understanding interactions between vectors and parasites/ pathogens and target based development of drug / insecticides.

ii) To assist the investigators to carry out genomic studies on parasites/pathogens/vectors of vector borne diseases

Advertisement: http://vcrc.res.in/Adv_Bio13.pdf

The Law group provides internal Institute-specific development, training and support roles for data manipulation, sequence analysis and any other aspect of the analysis of biological data using computer systems. Additionally we provide databases and applications supporting the international animal science community, particularly tools and resources for genome mapping.

Head: Andy Law. Members: John Bowman (animal facility database applications), Zen Lu (bioinformatics support), Trevor Paterson (software development)

More @ http://www.bioinformatics.ed.ac.uk/groups/roslin-bioinformatics-group