Team:CCU Taiwan/Notebook

•  CCU_Taiwan
•  Team
  Members Attribution Collaboration Sponsors
•  Project
  Description Overview Design Model Results Demonstration Safety Notebook Protocol
•  Parts
  Overview Basic Parts Composite Parts
•  Human Practices
  Integrated Human Practices Education & Engagement Entrepreneurship
•  Device
  Hardware Software
•  Medal Criteria
  Judging Form

LbCas12a protein crRNA Conjugation of ssDNA
with magnetic beads

crRNA

8/1~8/17 Cloning crRNAs into psB1A3 vector

8/20~8/27 Cloning DNA template of crRNAs into yTA vector.

8/30~9/11 PCR amplification of DNA template of crRNA for IVT assay

9/12~9/15 Cut positive control and negative control plasmid into linear

10/12~10/15 PCR amplification of DNA template of crRNA for IVT assay and purify crRNA

LbCas12a protein

7/7~7/9 Amplification and confirmation of plasmid encoded LbCas12a protein

7/22~7/29 Expression and confirmation of LbCas12a protein at 37℃.

7/30~8/4 Expression and confirmation of LbCas12a protein at 16℃

8/9~8/11 Tobacco Etch Virus (TEV) Protease mediated elution of LbCas12a protein

8/12~8/23 Prepare LbCas12a protein for FPLC purification

8/24~8/25 Examine the elution of LbCas12a protein in FPLC by SDS-PAGE analysis

8/26~9/6 Examine TEV enzyme digestion of LbCas12a protein

9/19~9/21 Improve the LbCas12a protein extraction

9/23~9/26 Optimize the TEV efficiency

10/15 Produce ssDNA from negative control plasmid

10/03~10/18 LbCas12a protein cis and trans cleavage

Conjugation of ssDNA with magnetic beads

10/20 Binding of ssDNA to magnetic beads