Related Experiment Video
Updated: Jun 17, 2026
A Universal Protocol for Large-scale gRNA Library Production from any DNA Source
Published on: December 6, 2017
[Construct cosmid libraries by isolating large genomic DNA fragments from Monascus ruber]
Yang An1, Jing Yang, Xinxin Xu
1Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. anyang@foxmail.com
Objective:
To isolate large genomic DNA fragments from Monascus ruber for the construction of cosmid libraries.
Methods:
Modified phenol-chloroform method was used to isolate genomic DNA. The isolated genomic DNA was digested by Sau3AI to 40kb fragments on average. Then, the fragments were packaged by Stratagene' s Gigapack III XL packaging extract. A pair of degenerate primers were used to amplify a fragment of PKS (polyketide synthase) gene from this cosmid library.
Results:
The average size of genomic DNA isolated by this method was larger than 48 kb, with a concentration of 5 microg/microl. The constructed cosmid libraries had 10 folders coverage of the Monascus spp. genome. A cosmid containing the homologue of PKS gene was obtained by PCR screening.
Conclusion:
This modified method of isolating large fragments genomic DNA of Monascus ruber was efficient and feasible.
