The three-year LGG fellowship training program has given me extensive experience in the analysis, interpretation and reporting of several clinical tests. I was also involved in test development, clinical research projects, quality improvement projects, and presentation of several projects at local and national level meetings. Clinical tests that I have actively participated in include: • Chromosome, FISH, and microarray analysis for cancer and constitutional disorders (pre- and postnatal)• Sanger sequencing, PCR fragment analysis, Methylation-Specific Multiplex Ligation-Dependent Probe Amplification (MS-MLPA)• Next-generation sequencing (NGS) tests including symptom-driven whole exome sequencing, disease gene panel analysis, and whole genome sequencing (for cancer and constitutional disorders)

As a new and first hire for this position in the Cytogenetics lab, my general job description was to help with mate-pair data analysis. With the expertise I have in primer design and PCR amplification, I got involved in developing a system for designing primers to confirm the chromosomal rearrangements identified by mate-pair sequencing analysis. I developed a standard operating procedure (SOP) for this process for the first version of mate-pair diagnostic testing that is now available from Mayo Clinic. I was also involved in various research projects with several Cytogenetics lab directors, two of which have been presented as abstracts at the Cancer Genomics Consortium 2016.

As a new research fellow in Dr. Peter Harris’s lab, my pilot project was to analyze a few previously published, phenotypically positive Autosomal Dominant Polycystic Kidney Disease (ADPKD) families that do not show linkage to the known ADPKD genes, PKD1 or PKD2. In the attempt to exclude PKD1/PKD2 linkage in these families, we discovered PKD1/PKD2 mutation in four out of five of these families (Paul BM et al, Kidney International, 2014) through Sanger sequencing. As a continuation of this project, I worked on characterizing a large cohort of mutation negative ADPKD families using whole exome sequencing (WES). We identified a third causative gene (GANAB) in both autosomal dominant polycystic kidney and liver disease through WES analysis- Porath B and Gainullin V et al, American Journal of Human Genetics, 2016. My second major project was to characterize a set of novel and indeterminate PKD1 variants found in some ADPKD families, using various in silico and in vitro tools. For this project, I designed plasmids with the desired nucleotide change using site directed mutagenesis and transfected these plasmids into HEK cells. I used the protein lysate obtained from these cell cultures for western blotting, localization and glycosylation experiments to look for any variation in the protein compared to its wildtype counterpart. I was also a part of the Genetics core in the Mayo Translational Polycystic Kidney Disease Center (MTPC) where we worked on various developmental projects including generation and maintenance of databases for ADPKD mutations, variants found in ADPKD pseudogenes and also protocols used in the lab.

Graduate student, mentor: Dr. Greg Vanden HuevelAs a doctoral candidate in Dr. Greg Vanden Heuvel’s lab, I worked on generating and analyzing a mouse model to study ADPKD (Paul BM et al, Developmental Dynamics, 2008). Our lab’s main research focus was on Cux1, a homeobox gene important during kidney development. Cux1 is ectopically expressed in the mouse models of PKD as well as in human ADPKD. The overall aim of my study was to elucidate the role of Cux1 in PKD. My thesis work was to test the hypothesis that Cux1 is required to develop PKD in an ADPKD mouse model. Using various mouse genetics, molecular biology, histology and physiological approaches, I was able to show that Cux1 haploinsufficiency results in the amelioration of PKD in our ADPKD mouse model. I also had the privilege to publish an invited review with Dr. Vanden Heuvel as a result of my thesis work (Paul BM and Vanden Heuvel GB, Wiley Interdisciplinary Reviews: Developmental Biology, 2014).

I worked in the Genetics lab of a hospital with specialization in infertility treatments. Patients were referred to the genetics lab either from the infertility clinic or from peripheral health care centers for suspected chromosomal anomalies or infertility problems. The routine work included karyotyping, PCR for Y- chromosome microdeletion and semen analysis. Following were the responsibilities and the specific techniques performed.Taking case history from patients and maintaining patient filesPhlebotomy techniquesSetting up cultures using blood, amniotic fluid, chorionic villus, and abortus materialSlide preparation and GTG (G bands by Trypsin using Giemsa) banding for karyotypingDetection of Y- chromosome microdeletion through PCR and agarose gel electrophoresisModification of karyogram using automated karyotyping softwareWriting research papersOrganizing state-level seminarsOrganizing genetic counseling sessionsPresentation of seminars related to geneticsPreparation of semen for Intra Uterine Insemination (IUI) and In- vitro Fertilization (IVF)Cryo preservation of semen sample for IUI and IVF