Team:Botchan Lab Tokyo/Parts notebook

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RESULTS LABORATORY NOTEBOOK MODELING

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Experiment Plan

At first,we inserted gfpgene into pSB1C3(first step)

Second,we inserted nifBpromoter into upstream of gfpgene in pSB1C3-gfp plasmid.

Lab notebook

9/10

PCR

sample

①pSB1C3　②pnifB

9/12

Electrophoresis and Gel extraction

sample

PCR product

9/13

Electrophoresis

sample

Extracted PCR product

9/14

In Fusion Cloning

pSB1C3+nifB

Transformation

9/17

PCR

sample

①pSB1C3+pnifB　②gfp

Electrophoresis

Gel Extraction

Preculture

transformant

9/18

Plasmid extraction of transformant

Electrophoresis of extracted plasmid

PCR for step 2

sample

pnifB

9/19

PCR

sample

①pSB1C3+gfp

Electrophoresis

sample

PCR product

minipreped product

transformant

9/20

Electrophoresis

sample

PCR product

PCR

sample

pSB1C3+gfp

9/23

PCR

sample

pSB1C3+gfp

9/24

Electrophoresis

sample

PCR product

Gel Extraction

9/25

In Fusion Cloning

sample

pnifB+pSB1C3-gfp

9/26

Transformation

sample

pnifB+pSB1C3-gfp

Insert check PCR

sample

transformant

9/27

Electrophoresis

sample

PCR product

9/28

PCR

sample

①pnifB+gfp

②pSB1C3+gfp

Electrophoresis

sample

PCR product

Transformation

Insert check PCR

sample

transformant

10/1

PCR

sample

pnifB

Miniprep

sample

①pSb1C3+gfp

②pSb1C3+pnifB+gfp

Electrophoresis

sample

minipreped product

Restriction digest

sample

①pSb1C3+gfp

②pSb1C3pnifBgfp

Electrophoresis

sample

restriction digested product

minipreped product

Fluorescence measurement

sample

transformant

10/2

Preparation of miniprep

10/3

Miniprep

sample

pSb1C3pnifBgfp

Measurement of fluorescence intensity

sample

pSb1C3+pnifB+gfp

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