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We received the sequence of S. thermophilus DGCC7710 CRISPR1 locus, which was graciously provided by Rodolphe Barrangou and Danisco, Inc. We also signed a nondisclosure agreement with said party. Promptly after the sequence was received we designed primers to amplify the entire locus.

Cultured cells overnight and performed a genomic DNA extraction. We used primers to amplify out the CRISPR locus via PCR.

Using the native plasmid pNT1 as a shuttling vector for the CRISPR was extremely ineffective, and we consistently got low yields of plasmid DNA.

During the adviser meeting, one of the important ideas in our CRISPR project is suggested by Dr. Hammer. It is found that bacillus subtilis which is a gram positive bacteria do not have the CRISPR mechanism present in them. Although, it is suggested that similar promoters might be present in B. subtilis as present in the Streptococcus thermophilus. So, we designed the experiments this week for introducing the CRISPR system in the bacillus subtilis1st round of Bacillus cultures arrive, are grown on LB media and agar.