# Epigenetic control of meiotic recombination in mammals - Equipment Supplement

> **NIH NIH R01** · OKLAHOMA MEDICAL RESEARCH FOUNDATION · 2021 · $129,704

## Abstract

Summary
Summary of parent project: The parent grant (R01GM125803) for this equipment request is entitled “Epigenetic
Control of Meiotic Recombination in Mammals”. This award covers the primary research focus of the laboratory
analyzing the interplay between chromatin-based control of DNA repair and the mechanics of chromosome
movement in mouse meiosis. A large percentage of our research productivity is achieved through generation of
genome wide maps of chromatin binding sites for our proteins of interest in wild type and germ cell genetically
engineered mice. This includes, purification of spermatocyte cells, and chromatin immunoprecipitation followed
by deep sequencing (ChIP-seq) and RNA-seq. This is complemented with observation of histological and cell
biology microscopic analysis of testis specific knockout mice for chromatin remodelers in mice spermatocytes.
Obtaining and analyzing genome wide profiles of protein chromatin binding sites using ChIP-seq. Generation of
ChIP-seq profiles: Regarding generation of DNA material necessary for identification of protein binding sites, we
often employ enriched fractions of mouse spermatocytes at different stages to obtain crosslinked DNA
fragments-protein complexes that allow us (after immunoprecipitation and DNA sequencing) assessing the
precise localization of protein of interest and how their distribution changes in specific mouse genetic
backgrounds and biochemical manipulations. This approach has shown to be very useful in studying chromatin
remodeling complexes composition and specificity of subunit functions, which are direct measurements of
chromatin remodeler activity on chromatin conformation and DNA repair.
Current instrumentation and equipment needed. Our laboratory confronts technical challenges due to the limited
accessibility and quality of the existent instrumentation (detailed below) employed to prepare DNA-protein
adducts and preparation of DNA libraries previous to DNA deep sequencing.
ChIP-seq processing and required instrumentation: After enriched fraction of mouse spermatocytes are obtained,
DNA-protein complexes are fixed with paraformaldehyde and DNA must be precisely shredded, in a specific
DNA range size that we previously determined, using a focused ultrasonicator (request Covaris E220 evolution).
At this stage of the procedure, is critical that DNA size and quality is carefully measured to evaluate the efficiency
of previous steps and allow the generation of suitable DNA libraries for sequencing. Quantity and quality
measurement require the use of at least two alternative measures. It is here that Microvolume UV-Vis
Spectrophotometer and Fluorometer measurement (request Qubit flex) of small volumes together allow you to
obtain the most complete information about the concentration and quality DNA samples. This is essential to
prevent costly troubleshooting downstream. In the next step, DNA-protein samples undergo immunoprecipitation
with specific antibodies and protein-DNA cr...

## Key facts

- **NIH application ID:** 10375710
- **Project number:** 3R01GM125803-04S1
- **Recipient organization:** OKLAHOMA MEDICAL RESEARCH FOUNDATION
- **Principal Investigator:** Roberto Jose Pezza
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $129,704
- **Award type:** 3
- **Project period:** 2018-07-05 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10375710

## Citation

> US National Institutes of Health, RePORTER application 10375710, Epigenetic control of meiotic recombination in mammals - Equipment Supplement (3R01GM125803-04S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10375710. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*