Improved protocols for BAC insert DNA isolation, BAC end sequencing and FISH for construction of BAC based physical map of genes on the chromosomes
| Autor: | Vijay Kumar Singh, Sathiya Murali, Vishwamitra Singh Baisvar, Basdeo Kushwaha, Ravindra Kumar |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Chromosomes Artificial Bacterial Positional cloning Computational biology Biology DNA sequencing Insert (molecular biology) 03 medical and health sciences 0302 clinical medicine Genetics Molecular Biology Gene In Situ Hybridization Fluorescence Genomic organization Gene Library Bacterial artificial chromosome Hybridization probe Computational Biology General Medicine DNA Sequence Analysis DNA Physical Chromosome Mapping DNA extraction 030104 developmental biology 030220 oncology & carcinogenesis Software |
| Zdroj: | Molecular biology reports. 47(3) |
| ISSN: | 1573-4978 |
| Popis: | Bacterial artificial chromosome (BAC) library is an important genomic resource useful in targeted marker development, positional cloning, physical mapping and a substrate for genome sequencing for better understanding the genome organization of a species. The present manuscript elucidates the improvement in protocols for economical and efficient BAC insert DNA isolation, BAC end sequencing and FISH for physical localization on the metaphase chromosome complements. BAC clones of Clarias magur, maintained in 384-well plate format in our laboratory, were used in this study. The protocols gave consistent and efficient results. We use routinely these protocols for BAC insert DNA extraction, generating end sequence data of the clone and constructing DNA probes to hybridize on the metaphase spreads of C. magur using FISH for physical their localization. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink