Difference between revisions of "Team:JNFLS/NoteBook"

 
Line 3: Line 3:
  
 
<div class="column full_size">
 
<div class="column full_size">
−
 
+
<br>
 +
<br>
 +
<br>
 
<p>    </p>
 
<p>    </p>
 
<h4>Week 1(4.9-4.15)</h4>
 
<h4>Week 1(4.9-4.15)</h4>

Latest revision as of 23:13, 17 October 2018




Week 1(4.9-4.15)

  • Recruitment of team members and first meeting.
  • Brainstorming and learning about relevant materials.
  • We started to discuss our ideas and invited teachers to give some advice about our projects.
  • Presentation within the team to share what we learn about synthetic biology.

    • Week2(4.23-4.29)

  • Finally, we choose the idea of creating a new detection method for HCV in donated blood.
  • A lab was provided by Shandong University in summer vacation.
  • After many discussions, everyone's division of labor is becoming clearer.
  • We began to use our spare time to study experimental techniques.
  • we started to brainstorm in order to create our team logo.

    • Week3 (5.13-5.20)

  • We had a safety education before we started the formal experiment, including lab safety, researcher safety and environment safety. And we learned some emergency responses, such as how to use extinguisher and fire blanket, and how to use emergency shower and eye wash.
  • Made a concrete plan for our project.

    • Week4 (5.21-5.27)

  • Preparing the pCMV-CE1E2 plasmid
  • Learning bacteria culture techniques, and beginning to culture E.Coli
  • We ordered primers for the PCR of HCVC gene extraction from Biomics

    • Week5 (5.28-6.3)

  • We received the primers
  • The PCR to extract HCVC gene from pCMV-CE1E2
  • HCVC gene was obtained from PCR, and we sent it to the company for sequencing
  • We chose and ordered pColdII plasmids from the company
  • We ordered endonuclease KpnI and PstI from the company

    • Week6 (6.4-6.10)

  • We received pColdII plasmids
  • Digestion of pColdII with KpnI and PstI
  • Digestion of HCVC gene with KpnI and PstI
  • Ligation of the plasmid and HCVC gene

    • Week7 (6.11-6.17)

  • Transformation of the plasmid containing HCVC gene, monoclonal selection, and culturing transformed E.Coli
  • Positive colony PCR, and positive clone selection
  • We went to local schools to have teaching activities for a week to spread IGEM and biology to children
  • Preparing of experimental materials.

    • Week8 (6.18-6.24)

  • SDS-PAGE for HCVC protein gained in the first batch
  • According to our topic, we designed a series of questionnaires.

    • Week9 (6.25-7.1)

  • After browsing references, we decided to truncate the HCVC gene into HCVC120 and HCVC173
  • Through a small range of issuing questionnaires, we have further modified our questionnaires.
  • We ordered primers for truncation

    • Week10 (7.2-7.8)

  • We received the primers
  • We went to Blood Center of Shandong to have a meeting with the researchers who are familiar with pathogen detection in blood samples
  • Truncating HCVC gene using PCR technology
  • Sending HCVC120 and HCVC173 to the company for sequencing
  • We started issuing questionnaires in a wide range.

    • Week11 (7.9-7.15)

  • Ligating HCVC120 and HCVC173 into vector plasmid pColdII.
  • Preparing for the work of InterLab.
  • We went to the local blood donation bus to make propaganda of our project and blood donation, and helped the doctors with blood donation
  • Transforming the plasmid into E.Coli and perform positive colony PCR
  • SDS-PAGE of the extracted protein expressed from HCVC120 and HCVC173
  • We decided to optimize the sequence of HCVC120 and HCVC173

    • Week12 (7.16-7.22)

  • Optimizing the sequence of HCVC120 and HCVC173
  • Continuing to Construct the plasmid vector.
  • For the InterLab, we have carried out preliminary experiments to be familiar with the use of instruments.
  • We went to the local book store to hand out our questionnaires and get the public informed of our project and synthetic biology
  • We obtained optimized sequence HCVCO120 and HCVCO173

    • Week13 (7.23-7.29)

  • We ligated HCVCO120 and HCVCO173 into the pColdII plasmid
  • We have done the work of InterLab
  • Transforming the plasmid into E.Coli, inducing expression, and collecting purified protein

    • Week14 (7.30-8.5)

  • SDS-PAGE of purified protein HCVCO120 and HCVCO173
  • We used 293T cell to further purify the protein
  • Learning cell culturing and transfection technology.
  • We have interview clinicians to know more about HCV and make some adjustments on our experimental ideas.

    • Week15 (8.6-8.12)

  • SDS-PAGE of purified protein(from 293T)
  • The HCVC protein was finally purified successfully after hard working; we have adequate amount of antigen to perform further experiment now
  • We have made our poster.

    • Week16 (8.13-8.19)

  • We ordered aptamers, padlock probes, and cDNAs needed in our system
  • Public propaganda of knowledge about blood donation in other local blood donation buses with the team members’ parents

    • Week17 (8.20-8.26)

  • We received the synthesized DNAs.
  • We participated in the CCiC and acquired much valuable experience.

    • Week18 (8.27-9.2)

  • We took part in the CCiC in Shanghai and acquired valuable advice.
  • We started to perform the experiments to validify the operability of our detection system, using the DNAs and HCVC proteins
  • Extracting the genomic DNA from cells, and starting exploring the best conditions of PCR.
  • It was validified that our detection system is operable.

    • Week19 (9.3-9.9)

  • We have launched a series of activities to promote the IGEM competition for Freshmen.
  • A presentation for the students in our own school

    • Week20 (9.10-9.16)

  • Further experiment on our aptamer-based detection system to find out the best condition for the detection.
  • Build parts, including original HCVC-protein-encoded gene, truncated HCVC-protein-encoded gene, codon-optimized HCVC-protein-encoded gene, and further optimized HCVC-protein-encoded gene.
  • We started writing wiki.

    • Week21 (9.17-9.23)

  • We discovered the most favorable condition for the detection to perform
  • Seven team members were select to join the Giant Jamboree.
  • We started to measure the binding rate of HCVC protein and aptamer at different aptamer concentrations over time.

    • Week22 (9.24-9.30)

  • Starting to analysis the data we have monitored, and looking for help in modeling.
  • Continuing to measure the binding rate of HCVC protein and aptamer at different aptamer concentrations over time.

    • Week23 (10.1-10.7)

  • Sorting out experimental ideas and establishing correlation model.
  • Continuing writing Wiki

    • Week24 (10.8-10.14)

  • After data processing, we got the curve which showed the effects of different concentrations of HCVC7 aptamer on the result of HCVC protein detection in our system
  • We have designed our winter uniforms.
  • Continuing writing Wiki

    • Week25 (10.15-10.21)

  • We have sent all of our parts to the Genscript, Nanjing, China.