Title Page
Abstract
Contents
Ⅰ. Introduction 10
Ⅱ. Materials and Methods 12
2.1. Zebrafish 12
2.2. Cloning of random barcode libraries 12
2.3. Preparation of embryos from various developmental stages 13
2.4. Barcoded embryo generation and Dissociation 13
2.5. cDNA analysis from barcode injected embryo 13
2.6. FACS sorting of barcoded cells 13
2.7. Quantitative PCR with barcoded cells 14
Ⅲ. Results 15
3.1. Designing random barcode for genomic integration 15
3.2. Random Barcode integration to zebrafish genome 16
3.4. FACS sorting of Barcoded-Embryos 17
3.5. Examining neural crest markers from random barcoded cells 17
Ⅳ. Discussion 33
Ⅴ. Abstract in Korean (국문 초록) 36
Ⅵ. References 37
Figure 1. Schematic diagram of overall experimental flow 19
Figure 2. Designing and cloning random barcoded plasmid. 24
Figure 3. Random barcode injection into wild-type embryos. 25
Figure 4. FACS sorting and qPCR of random barcoded transgenic zebrafish. 29
Supplementary Figure 1. Fluorescence microscope images of random-barcode injected AB embryos (24hpf) 27
Supplementary Figure 2. Fluorescence microscope images of random-barcode injected Tg(Sox10:Gal4; UAS:EGFP) embryos of various developmental stages. 30
Supplementary Figure 3. FACS result of control samples 32