RESEARCH SKILLS

Practice of handling experimental laboratory animals:

• • Perianal injection of drug/ microRNA/ siRNA targeting internal anal sphincter

  • Perianal injection of microRNA/ siRNA, followed by magentofection targeting delivery to internal anal sphincter

  • Rectoanal manometry to measure intraluminal pressure of rat internal anal sphincter

  • Ovarioectomy, implantation of osmotic pump with drugs, generating conditional knock down mice

Physiological Studies:

• • Handling Automatic Organ Bath

  • Isometric contraction study and electric field stimulation studies (using rat internal anal sphincter smooth muscle, rat mesenteric artery, caprine superior mesenteric artery, caprine iliac artery, caprine ruminal artery, caprine umblical artery, caprine uterine artery, caprine uterine muscle)

  • Pressure arteriography (Using Mesenteric Artery)

  • Pressure natriuresis and renal blood flow monitoring

Cell Biology:

• • Isolation and culture of circular smooth muscle cells from internal anal sphincter, rectum and colon of rodents and human

  • Isolation of endothelial cells from goat umbilical artery

  • Generation of bioengineered internal anal sphincter rings from circular smooth muscle cells

  • Morphometric analysis of smooth muscle cell for agonist and antagonist induced contraction study

  • siRNA transfection using Lipofectamine and Magnetofection to knockdown target genes

  • Mice embryonic fibroblast isolation, culture and maintenance

Molecular Biology:

• • Isolation of total RNA and cDNA synthesis

  • RT-PCR, Real Time PCR, genotyping

  • Agarose or polyacrylamide gel electrophoresis

  • Primer design for RT-PCR and qPCR

Protein Biochemistry:

• • Protein estimation and preparation: Organelle (nuclear) and cytosolic fractionations.

  • Protein expression and interaction: Western blot, immuno-precipitation and co-immuno-precipitation

  • Receptor biotinylation for tracking receptor internalisation and their recycling back

  • Sandwich ELISA

Fluorescence Microscopy:

• • Immunocytochemistry and immunohistochemistry with epifluroscence and confocal

  • Dihydroethidium staining to detect reactive oxygen species

Software Specific Skills:

• • Labchart version 8

  • GraphPad Prism 6

  • Bioinformatics tools: BLAST-protein and nucleotide, DAVID, KEGG, VARELECT, Gene Ontology

  • Microsoft Office, Excel, Power Point,

  • Adobe Photoshop, Image J (western blot and immunocytochemistry)

  • Ion Optix