Molecular combing in the analysis of developmentally regulated amplified segments of Bradysia hygida

• DOI: 10.4238/2012.august.6.10
• Publication Date: 2012-08-08T17:04:03Z
• Authors: K.J.R. Passos; S.Y. Togoro; S. Carignon; S. Koundrioukoff; A.-M. Lachages; M. Debatisse; M.A. Fernandez
• Keywords: Combing; Polytene chromosome; Amplicon

Molecular combing technology is an important new tool for the functional and physical mapping of genome segments. It designed to identify amplifications, microdeletions, rearrangements in a DNA sequence study process replication. This technique has recently been used analyze dynamics replication amplified domains. In Bradysia hygida, multiple amplification initiation sites are predicted exist upstream BhC4-1 gene. However, it impossible them using available standard techniques. The aim this was optimize molecular obtain fibers from polytene nuclei salivary glands B. hygida organism. Our results suggest that without prior purification possible. density, integrity, linearity were analyzed, 50 300 kb length detected, 9-kb fragment within region visualized biotin detected by Alexa Fluor 488-conjugated streptavidin technique. feasibility physically these demonstrated suggests may be origin amplicon.