Results
Results
Metagenomic Library
DNA Extraction We were able to confirm the size of the metagenomic DNA isolated from the porcupine fecal samples (Fig.1). The remainder of the DNA was ran via PFGE and all DNA from 24.8 kB was excised without exposure to UV or ethidium bromide. The gel was stained after words for visualization (Fig. 2). Vector Preparation Trouble-Shooting Future Directions of the cosmid librarySequencing Meagenomic and Cloning
Over the course of the 2017 iGEM season, we have had some downs, but many more ups.Project Achievements
Successes- Developed a pipeline to identify, or "mine", the porcupine metagenomic sequencing to discover novel enzymes.
- Identified 8? novel enzymes with variable percent identity.
- Synthesized 5 of those enzymes, and successfully cloned 4 of them into psB1AK3.
- Optimized our previous biobrick Endoglucanse(BBa_K2160000)by adding a C termincal HIS-tag and N terminal PelB sequence (Improve).
- Successfully completed a fluorophore cleavage assay from the Hallam lab.
- Isolated high molecular weight DNA from porcupine fecal samples.
- Obtained efficient ligation and digestion with pJC8 controls.
- Produced phage plaque with the phage packaging extract lamba DNA controls.
- Did not clone our biobricks into the shipping vector psB1C3
- Was not able to achieve the right environment for our novel beta-xylanase to function
- Was not able to design a functional media assay for our enzymes
- Could not make a functional metagenomic library
- Sheered our high molecular weight DNA