Notebook

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

Creating parts of HlyA and GFP

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 Ligation the GFP+HlyA in 9/18 was Electrophoresis

Date:9/19 DNA purify and PCR

Date:9/20 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:9/25

Date:9/26

Date:9/27

Date:9/28

Date:9/29

Date:9/30

Date:10/1

Date:10/2

Date:10/3

Date:10/4

Date:10/5

Date:10/6

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:9/1

Date:9/2 PCR and Electrophoresis

Date:9/3

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 Colony PCR and DNA purification and sequence

Date:10/6

Date:10/7

Date:10/8

Date:10/9

Date:10/10

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:9/1

Date:9/2

Date:9/3

Date:9/4

Date:9/5

Date:9/6

Date:9/7

Date:9/8

Date:9/9 Sequence(to To confirm whether variation happened in DNA of IL-10α,IL-10β)

Date:9/10-10/9 Sequence preparation & Sequence