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Yikon genomics Co. Ltd.

6 ARTICLES PUBLISHED IN JoVE

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Genetics

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format
Muhammad Khairul Ramlee 1, Jing Wang 1, Alice M. S. Cheung 1, Shang Li 1
1Cancer & Stem Cell Biology Programme, Duke-NUS Medical School

The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.

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Neuroscience

Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method
Thomas R. Groves 1,2,3, Jing Wang 1,2, Marjan Boerma 1,2, Antiño R. Allen 1,2,3
1Division of Radiation Health, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 3Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences

Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting.

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Non-invasive Chromosome Screening (NICS) of Human Preimplantation Embryos: Sample Collection and Chromosomal Ploidy Analysis by MALBAC-NGS
Rui Fang *1, Yaxin Yao *2, Yiyun Chen 2, Jieliang Ma 2, Li-Yi Cai 1, Sijia Lu *2
1Reproductive Medicine Centre, Wuxi Maternity and Child Health Hospital Affiliated to Nanjing Medical University, 2Department of Clinical Research, Yikon Genomics Co. Ltd.

We report a protocol for chromosome screening of human embryos by using spent culture medium, which avoids embryo biopsy and enables reporting chromosome ploidy using next generation sequencing (NGS). We present the detailed procedure including the preparation of culture medium, whole genome amplification (WGA), NGS library preparation, and data analysis.

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Neuroscience

Intracranial Pharmacotherapy and Pain Assays in Rodents
Erik Martinez 1, Haocheng Zhou 1, Jing Wang 1,2
1Department of Anesthesiology, Perioperative Care and Pain Medicine, New York University School of Medicine, 2Department of Neuroscience and Physiology, New York University School of Medicine

Here we present a protocol to perform intracranial pharmacological experiments followed by pain behavior assays in rodents. This protocol allows researchers to deliver molecular and cellular targets in the brain, for pharmacologic agents in the treatment of pain.

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Biology

Quantification of Proliferative and Dead Cells in Enteroids
Hua-Shan Li *1, Shao-Fang Xu *1, Jian-Ying Sheng *1, Zhi-Hui Jiang 1, Jing Wang 1, Ning Ding 1, Tao Wang 1, Matthew A. Odenwald 2, Jerrold R. Turner 2,3, Wei-Qi He 1, Hong Xu 1, Juan-Min Zha 1
1Jiangsu Key Laboratory of Neuropsychiatric Diseases and Cambridge-Suda (CAM-SU) Genomic Resource Center, Medical College of Soochow University, Department of Oncology, The First Affiliated Hospital of Soochow University, 2Department of Pathology, University of Chicago, 3Department of Pathology, Brigham and Women's Hospital–Harvard Medical School

The presented protocol uses flow cytometry to quantify the number of proliferating and dead cells in cultured mouse enteroids. This method is helpful to evaluate the effects of drug treatment on organoid proliferation and survival.

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Genetics

Chromosome Screening of Human Preimplantation Embryos by Using Spent Culture Medium: Sample Collection and Chromosomal Ploidy Analysis
Jin Huang *1,2,3,4, Yaxin Yao *5, Jialin Jia 1,2,3,4, Xiaohui Zhu 1,2,3,4, Jieliang Ma 5, Jing Wang 5, Ping Liu 1,2,3,4, Sijia Lu 5
1Centre for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, 2National Clinical Research Center for Obstetrics and Gynecology(Peking University Third Hospital), 3Ministry of Education, Key Laboratory of Assisted Reproduction (Peking University), 4Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, 5Department of Clinical Research, Yikon Genomics Co. Ltd.

The present study reports a protocol for chromosome screening of human embryos that uses spent culture medium, which avoids embryo biopsy and enables chromosome ploidy identification using NGS. The present article presents the detailed procedure, including the preparation of culture medium, whole genome amplification (WGA), next-generation sequencing (NGS) library preparation, and data analysis.

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