Gap closure with multiplex PCR and sequencing
The Multiplex PCR and Sequencing strategies were based on those described in: Optimized Multiplex PCR: Efficiently closing a whole-genome shotgun sequencing project. H. Tervelin, D. Radune, H. Khouri and S.L. Salzberg. (1999). Genomics 62, 500-507.
To this protocol, we brought few modifications to enhance the efficiency of the Multiplex strategy and to be able to close 100 gaps in only 3 steps...
Download here our detailed Multiplex protocol describing the strategy used for constructing the primers mixes