Team:NU Kazakhstan/Timeline

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May

This month was used for preparation of all equipment, media and reagents that can be necessary for realisation of the project. Also, in May newly recruited and future members of iGEM team acquired skills in basic techniques of biotechnology. The plan of the activities and protocols was worked out in order to keep the big picture of the process and organise the work of all part of the project.

Week 1

24.05.17

LB broth media and LB agar plates were prepared
Ampicillin stock was prepared using 1g of antibiotic in 10 ml of dH2O. Stock was stored in the fridge
Inoculation was performed in 15 ml Falcon tubes, using 5ml LB broth. Single colony was selected from LB agar, the tip was dropped and swirled. Bacterial culture was incubated at 37C in incubator

25.05.17

Miniprep was performed to extract pJD67 plasmid
Digestion of pJD67 plasmid was done
Gel electrophoresis was done using 1.5 % agarose gel, 120V

26.05.17

TAP media was prepared

27.05.17

Subculture was done by transferring 100 ul of algae to fresh TAP medium

28.05.17

Agar washing was done

29.05.17

OD of growing algae was measured
Agar washing was done

30.05.17

OD of growing algae was measured
Maxiprep was done to extract pJD67 plasmid
Agar washing was done

31.05.17

Ethanol precipitation (pJD67) was performed
TAE buffer, LB broth was prepared
Agar washing was done using ethanol
Inoculation of E.coli was done (5 ml of LB broth)

June

In June we focused on collection of the stock DNA which could be used further in electroporations. Miniprep extraction kit seemed to be unpersuasive in its concentrations and we tried to work with Maxiprep with a purpose of obtaining at least 2 ug of plasmid DNA for C.reinhardtii electroporation. At the same time, we used June to develop a biosafety system SuperNova for our project.

1.06.17

OD of growing algae was measured
Ampicillin stock was prepared

2.06.17

Transformation of E.coli was performed using pJD67
OD was measured

3.06.17

Inoculation of transformed cells was done
OD of growing algae was measured
Miniprep was done, concentration was measured
Gel electrophoresis was done using 1.5 % gel.

OD

Nanodrop results

5.06.17

Transformation of E.coli with pHyg3 and pJD67
Chromium media (0.2mM, 100mM) was prepared
Agar plates (arg+/hyg+, arg+/ hyg-, arg-/hyg+, arg-/hyg-) were prepared

6.06.17

Agar plates with ampicillin and amphotericin B were prepared
Competent cells were prepared

8.06.17

Transformation of E.coli using pJD67

10.06.17

Inoculation was done, from liquid media of algae to the plates with ampicillin+amphotericin B

15.06.17

Starch washing was done, starch was stored in ethanol (+4C)

16.06.17

Digestion of pHyg3 with HindIII was done

17.06.17

Inoculation was done for miniprep, to extract pJD67 and pHyg3 plasmids
Agar washing was done

18.06.17

Miniprep was done (both pJD67 and pHyg3)
Concentrations were measure, gel electrophoresis was done

19.06.17

Digestion of pJD67 was done using HindIII
Concentration was measured using Nanodrop

Digestion of pHyg3 with KpnI was done
Concentration was measured using Nanodrop

20.06.17

Transfection of algae with pHyg3 and pJD67 was done

21.06.17

Plating of transfected was done
Media was prepared

July

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