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From 2010.igem.org

Notebook	Building Parts	Testing Parts	Assembly Method	Competent Cells	Plates	Kit Manual	Software

Plates

Sintered Plastic

22-06-10

Creation of a reusable surface on which to grow bacteria.

Supplier: SPC Technologies Ltd. Sintered Plastic Samples

1) 3.0mm Ultra-Fine PE Sheet - mean pore size 14µm

2) 6.0mm Fine Grade PE Sheet - mean pore size 30µm

3) 4.5mm Medium Grade PE Sheet - mean pore size 88µm

• according to materials sheet, the ultra-fine sheet is a bacterial barrier

Procedure

1) each 100mm x 100mm sample was cut into...

TO BE CONTINUED All of the following steps were performed in a safety cabinet under sterile conditions.

1) disassembled a Degree deodorant stick and soaked in ethanol

2) removed the raising platform and covered it with Parafilm

3) coated the insides of the tube with mineral oil

5) removed platform and poured LB agar into the base of the stick, waited until it solidified

6) the Parafilmed platform was put back into the stick and lowered maximally

7) LB agar was poured on top of the platform, until the tube was full, waited until it solidifed

8) The top of the LB agar was sliced off with an ethanol-sterilized knife

9) 400ul of transformed competent cells were plated on the stick as if it was a standard LB agar plate

10) L.B.O. stick was incubated overnight at 37C

Observations:

- although the knife cut through the LB agar easily, it was difficult to produce a completely flat surface

- solidified LB agar was effectively raised and lowered using the dial

- after incubation, a bacterial lawn was observed and a distinct E.coli smell was present

29-06-10

1) Followed procedure from June 26, 2010 to put new LB agar into the L.B.O. tube

2) Approximately 1cm was sliced off the top of the new LB agar with an ethanol-sterilized razor blade

3) L.B.O. stick was incubated overnight at 37C

Observations:

- no growth observed, tube must have been sterile

30-06-10

1) Streaked green colony from a plate of Cambridge parts

2) L.B.O. stick was incubated overnight at 37C

Observations:

- green streak and individual green colonies observed, along with a distinct E.coli smell

05-07-10

1) Approximately 1cm was sliced off the top of the used LB agar with an ethanol-sterilized razor blade

2) Streaked red, RFP-containing colony onto the LB agar surface.

3) L.B.O. stick was incubated overnight at 37C

Observations:

- only a red streak and individual red colonies observed, along with a distinct E.coli smell

- no remnants of green colonies visible

L.B.O.

24-06-10

Creation of "L.B.O.": a deodorant stick of LB agar

All of the following steps were performed in a safety cabinet under sterile conditions.

1) disassembled a Degree deodorant stick and soaked in ethanol

2) removed the raising platform and covered it with Parafilm

3) coated the insides of the tube with mineral oil

5) removed platform and poured LB agar into the base of the stick, waited until it solidified

6) the Parafilmed platform was put back into the stick and lowered maximally

7) LB agar was poured on top of the platform, until the tube was full, waited until it solidifed

8) The top of the LB agar was sliced off with an ethanol-sterilized knife

9) 400ul of transformed competent cells were plated on the stick as if it was a standard LB agar plate

10) L.B.O. stick was incubated overnight at 37C

Observations:

- although the knife cut through the LB agar easily, it was difficult to produce a completely flat surface

- solidified LB agar was effectively raised and lowered using the dial

- after incubation, a bacterial lawn was observed and a distinct E.coli smell was present

29-06-10

1) Followed procedure from June 26, 2010 to put new LB agar into the L.B.O. tube

2) Approximately 1cm was sliced off the top of the new LB agar with an ethanol-sterilized razor blade

3) L.B.O. stick was incubated overnight at 37C

Observations:

- no growth observed, tube must have been sterile

30-06-10

1) Streaked green colony from a plate of Cambridge parts

2) L.B.O. stick was incubated overnight at 37C

Observations:

- green streak and individual green colonies observed, along with a distinct E.coli smell

05-07-10

1) Approximately 1cm was sliced off the top of the used LB agar with an ethanol-sterilized razor blade

2) Streaked red, RFP-containing colony onto the LB agar surface.

3) L.B.O. stick was incubated overnight at 37C

Observations:

- only a red streak and individual red colonies observed, along with a distinct E.coli smell

- no remnants of green colonies visible