Team:KAIT Japan/Notebook

From 2014.igem.org

Revision as of 09:39, 17 October 2014 by Iguana (Talk | contribs)
ロゴ2.png

KAIT Japan2013 Kanako.png

KAIT Japan 2014 iGEMRogo.png
New home t.jpg

Home

New Team1.jpg

Team

New 考える人.jpg

Project

New 歯車.jpg

Parts

New 111111111111111.jpg

Protocol

Ff.jpg

Notebook

New 結果.jpg

Results

New new 危険.jpg

Safety

New new ひと.jpg

Human Practice


Notebook

We made an experiment every day. It was process at trial and error.



Creating parts of GFP and HlyA

Date:8/22 The refinement of DNA and PCR and Electrophoresis
Date:8/25 PCR and Electrophoresis and Restriction
Date:8/26 Insert gene into TAvectar and Transformation
Date:8/26 Blue white Selection and Electrophoresis(8/25 Restriction) and DNA Extraction
Date:8/27 Electrophoresed the DNA that We did refinement on August 26 to confirmed it
Date:8/28 Check the HlyA and GFP for colony PCR
Date:8/29 DNA purify the HlyA and GFP
Date:8/29 Ligation product and check for PCR
Date:9/1 Build replica the HlyA and GFP
Date:9/1 Restriction enzyme treatment the HlyA and GFP
Date:9/2 Restriction enzyme treatment the HlyA and GFP in 9/1 was NANOSEP
Date:9/3 Ligation and PCR, DNA purification the HlyA and GFP
Date:9/4 Restriction enzyme treatment the HlyA and GFP
Date:9/5 Restriction enzyme treatment the vector
Date:9/5 Ligation the vector and insert. So transformation.
Date:9/9 Check the HiyA and GFP replica.Ligation and transformation the vector and insert DNA.
Date:9/10 Transformation in 9/9 check the colony PCR.TA cloning the HlyA+GFP.
Date:9/11 Ligation the vector and insert DNA.
Date:9/12~9/15 Transformation the Ligation product.
Date:9/16 Noticed wrong the primer.so restart from the First.
Date:9/17 PCR the DNA abstracted for iGEM kit.It was TA cloning and restriction enzyme treatment.
Date:9/18 Ligation the GFP and HlyA.
Date:9/19 Electrophoresis the ligation the GFP+HlyA in 9/18
Date:9/20 DNA purify and PCR
Date:9/21 PCR and DNA purify
Date:9/22 Ligation the GFP+HlyA
Date:9/23 PCR and DNA purify
Date:9/24 TA Cloning was GFP and HlyA
Date:9/26 Restriction enzyme treatment the ligation the GFP+HlyA in 9/22
Date:9/27 Check the TA cloning of GFP+HlyA
Date:9/29 Restriction enzyme treatment the GFP+HlyA and the pSBIC3
Date:9/29 The vector and the insert were NANOSEP and lectrophoresis
Date:9/30 The vector and the insert were ligation and transformation
Date:9/30 The GFP+HlyA was PCR and DNA purify
Date:10/1 Electrophoresis the GFP+HlyA was PCR and DNA purify in 9/30
Date:10/2 Restriction enzyme treatment and ligation the GFP+HlyA
Date:10/3 Check the ligation in 10/2
Date:10/3 Transformation the plasmid
Date:10/5 Colony PCR
Date:10/6 Miniprep and Electrophoresis




Creating parts of STAT3

Date:8/19 PCR(8/18 Miniprep) and Electrophoresis
Date:8/20~8/22 PCR(Lowered 2℃ from Tm value.) and Electrophoresis
Date:8/25 PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
Date:8/26 PCR and Electrophoresis(Using a mutation primer.)
Date:8/27 First Variation introduction (with Prime STAR Max)
Date:9/2 PCR and Electrophoresis
Date:9/5~9/8 PCR and Electrophoresis
Date:9/9 Sequence(to To confirm whether variation happened in DNA of STAT3 )
Date:9/10~9/12 PCR and Electrophoresis
Date:9/13~9/15 DNA purification and electrophoresis
Date:9/16~9/17 DNA purification and colony PCR(Failure)
Date:9/18 colony PCR(Failure)
Date:9/19~9/22 colony PCR and electrophoresis
Date:9/23 PCR and Electrophoresis(We found out that DNA polymerase was malfunction)
Date:9/24 PCR and Electrophoresis and DNA DNA purification
Date:9/25 PCR(to find annealing temperature)
Date:9/26~9/30 DNA purification and electrophoresis(to use DNA for the sequence)
Date:10/1 Dilution after Concentration measurement of DNA
Date:10/2 Sequence(to check mutation of DNA)/The variation was not found.
Date:10/4 PCR and electrophoresis
Date:10/5~10/7 Colony PCR and DNA purification
Date:10/8 Concentration measurement of DNA
Date:10/9 Sequence(to check mutation of DNA)/The variation was not found




Creating parts of IL-10α,IL-10β

Date:8/19 PCR(8/18 Miniprep) and Electrophoresis
Date:8/20~8/22 PCR(Lowered 2℃ from Tm value.) and Electrophoresis
Date:8/25 PCR and Electrophoresis(Changed to Taq HOTSTER from EX taq.)
Date:8/26 PCR and Electrophoresis(Using a mutation primer.)
Date:8/27 First Variation introduction (with Prime STAR Max)
Date:9/2 PCR
Date:9/4 Transformation
Date:9/5 Colony PCR
Date:9/7 ColonyPCR
Date:9/8 DNA extraction and Sequence(to To confirm whether variation happened in DNA of IL-10β)
Date:9/9 DNA sequence
Date:9/10 PCR and Electtrophoresis
Date:9/11 Colony PCR
Date:9/12 DNA extraction
Date:9/13 PCR and Electrophoresis
Date:9/15 ultrafiltration
Date:9/16 DNA sequence(to comfirm whether variation happened in DNA of IL-10β)
Date:9/20 PCR
Date:9/23 Colony PCR
Date:9/24 Electrophoresis
Date:9/26 ultrafiltration
Date:9/27 PCR and colony PCR
Date:9/29 Electrophoresis
Date:9/30 DNA Extraction
Date:10/1 Colony PCR and DNA extraction
Date:10/2 Electrophoresis
Date:10/3 PCR(using Prime Star MAX Premix) and Electrophoresis
Date:10/4 PCR(using Prime Star MAX Premix) and Electrophoresis
Date:10/5 Electrophoresis



Creating parts of HRV,IL-5Ra,IL-5Rb,AraC,Arac Promoter

Date:9/16 PCR:Ara,AraPro,HRV Transformation:IL-5Ra,IL-5Rb
Date:9/17 Restriction enzyme treatment and insert to TA vector and transformation
Date:9/18 transformation
Date:9/19 miniprep and colony PCR
Date:9/20~9/22 PCR
Date:9/23 TA Cloning
Date:9/24 TA Cloning and PCR
Date:9/25 Restriction enzyme treatment and PCR
Date:9/26 Colony PCR and Ligation
Date:9/27 TA Cloning and Ligation and colony PCR
Date:9/28 PCR
Date:9/29 colony PCR and PCR and DNA refinement and Ligation
Date:9/30 DNA refinement and Ligation
Date:10/1 Restriction enzyme treatment and Ligation
Date:10/2 PCR and Ligation and DNA refinement
Date:10/3 Ligation
Date:10/4 Ligation
Date:10/7 PCR
Date:10/8 PCR and Restriction enzyme
Date:10/9 DNA refinement and Restriction enzyme
Date:10/10 Phoshatase treatment
Date:10/11 DNA refinement and Ligation and Restriction enzyme treatment
Date:10/12 DNA refinement and PCR and Ligation and phoshatase treatment