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

Date:9/21

Date:9/22

Date:9/23

Date:9/24

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

Date:9/14

Date:9/15

Date:9/16

Date:9/17

Date:9/18

Date:9/19

Date:9/20

Date:9/21

Date:9/22

Date:9/23

Date:9/24

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 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