Team:KAIT Japan/Notebook

Notebook

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