Team:Gunma/Notebook

NOTEBOOK

~May

Training Experiments

June

10th-19th
Planning Experiments
Searching for Information

20th
Culture BL21(DE3)pLysS in LB medium and measure OD600
21th
Culture BL21(DE3)pLysS in NaCl sol. of a variety of concentrations and measure OD600

August

23th
Culture BL21(DE3)pLysS in LB medium and measure OD600
Culture BL21(DE3)pLysS in NaCl sol. of a variety of concentrations and measure OD600
25th
Searching for Information
29th
Transform mag-2 pUBnew to BL21(DE3)pLysS,culture
Checked if BL21(DE3)pLysS can grow or not

September

13th
Transform MPX-G to BL21(DE3)
14th
Design primers for PCR of mag-2 and MPX plasmids
Preparation of 100 mM IPTG stock
Looking for the proper concentration of IPTG

17-18th
Transform MPX to BL21(DE3)pLysS,culture,add IPTG
Measure OD600
21th
PCR mag-2 and MPX sequense
Agarose electro phoresis
25th
PCR mag-2,MPX

26th
Preparation of 10X TBE buffer
27th
Agarose electro phoresis
29th
Treat samples with cloning enhancer
In-Fusion
Transformation
Culture overnight

October

1st
Plasmid extraction of mag-2 DH5-alpha,MPX DH5-alpha
2nd
Culture mag-2 DH5-alpha,MPX DH5-alpha

4th
Design primers to modify mag-2 DNA to combine with linearized BBa_I746909 on pSB1C3
Looking for the proper concentration of IPTG

5th-9th
Design and order primers

8th
Transformation of RFP pSB1C3 into DH5-alpha
Culture
9th
PCR

11-12th
Making Abs600 Standard curve
Making Fluorescent Standard curve

14th
Transformation of BBa_I746909 on pSB1C3 into BL21(DE3)

16th
Plasmid extraction of BBa_I746909 on pSB1C3
PCR to modify mag-2 sequence
Inverse PCR of BBa_I746909 on pSB1C3 to linearize it
Cut pSB1A3 vector and BBa_K3050000 insert by EcoRI and PstI
Agarose electro phoresis
Ligation
Transformation into BL21(DE3)
Culture overnight