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Team:Alberta/Notebook/ReusablePlates
From 2010.igem.org
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{{Team:Alberta/beginMainContent}} | {{Team:Alberta/beginMainContent}} | ||
=Creating Inexpensive and Reusable Plates= | =Creating Inexpensive and Reusable Plates= | ||
| - | === | + | |
| + | ===Project Timeline: Click on an image to see more information=== | ||
<html> | <html> | ||
| - | <img src="https://static.igem.org/mediawiki/2010/0/09/PlatesTimeline.jpg"> | + | <img src="https://static.igem.org/mediawiki/2010/0/09/PlatesTimeline.jpg" border="0" usemap="#Plates"> |
| + | <map name="Plates"> | ||
| + | <area shape="rect" coords="0, 0, 100, 83" href="https://2010.igem.org/wiki/index.php?title=Team:Alberta/Notebook/ReusablePlates#Scintered_Plastic_Plates"> | ||
| + | <area shape="rect" coords="191, 0, 290, 82" href="https://2010.igem.org/wiki/index.php?title=Team:Alberta/Notebook/ReusablePlates#L.B.O."> | ||
| + | <area shape="rect" coords="367, 25, 466, 72" href="https://2010.igem.org/wiki/index.php?title=Team:Alberta/Notebook/ReusablePlates#Jello_and_Gelatine_Plates"> | ||
| + | </map> | ||
</html> | </html> | ||
| - | + | ---------- | |
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| + | The equipment and supplies required to sterilize and make plates are expensive and not usually available to a high school lab. We attempted to create plates which could easily be sterilized by a microwave or bleach. | ||
==Scintered Plastic Plates== | ==Scintered Plastic Plates== | ||
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Scintered Plastic Supplier: SPC technologies Ltd. | Scintered Plastic Supplier: SPC technologies Ltd. | ||
| - | We used 3 | + | We used 3 types of scintered plastic: |
:*3.0mm Ultra-Fine PE sheet(14um pore-size) | :*3.0mm Ultra-Fine PE sheet(14um pore-size) | ||
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# 400ul of transformed competent cells were plated on the stick as if it was a standard LB agar plate | # 400ul of transformed competent cells were plated on the stick as if it was a standard LB agar plate | ||
# L.B.O. stick was incubated overnight at 37C | # L.B.O. stick was incubated overnight at 37C | ||
| + | <html> | ||
| + | <center> | ||
| + | <img src="http://farm5.static.flickr.com/4103/5116068476_a7c90747ba_m.jpg" width="240" height="180" alt="LBO" /> | ||
| + | </center> | ||
| + | </html> | ||
Observations: | Observations: | ||
:* although the knife cut through the LB agar easily, it was difficult to produce a completely flat surface | :* although the knife cut through the LB agar easily, it was difficult to produce a completely flat surface | ||
Latest revision as of 03:59, 28 October 2010
Creating Inexpensive and Reusable Plates
Project Timeline: Click on an image to see more information
The equipment and supplies required to sterilize and make plates are expensive and not usually available to a high school lab. We attempted to create plates which could easily be sterilized by a microwave or bleach.
Scintered Plastic Plates
Scintered Plastic Supplier: SPC technologies Ltd.
We used 3 types of scintered plastic:
- 3.0mm Ultra-Fine PE sheet(14um pore-size)
- ultra-fine grade should be a bacterial barrier
- 6.0mm Fine-Grade PE Sheet(30um pore-size)
- fine grade should also be a bacterial barrier but it was too hard to cut
- 4.5mm Medium Grade PE Sheet(88um pore-size)
Procedure:
- tested plastic's ability to absorb LB
- June 22, 2010 - added 10ml of LB and 40ul chloramphenicol to plates made from the 3.0mm and 4.5mm scintered plastics. Incubated in a 37C warm room overnight.
- June 23, 2010 - 7ml LB left in 3.0mm ultra-fine and 8ml left in 4.5mm medium-grade
- Streaking on Plastic
- June 23, 2010 - streaked both types of scintered plastic with RFP-containing colonies(after transferring plastic into new petri dishes). Left in 37C warm room overnight.
- June 24, 2010 - no growth after incubation, plastic was very dry
L.B.O
Creation of "L.B.O.": a deodorant stick of LB agar
- June 24, 2010 - All of the following steps were performed in a safety cabinet under sterile conditions.
- disassembled a Degree deodorant stick and soaked in ethanol
- removed the raising platform and covered it with Parafilm
- coated the insides of the tube with mineral oil
- removed platform and poured LB agar into the base of the stick, waited until it solidified
- the Parafilmed platform was put back into the stick and lowered maximally
- LB agar was poured on top of the platform, until the tube was full, waited until it solidifed
- The top of the LB agar was sliced off with an ethanol-sterilized knife
- 400ul of transformed competent cells were plated on the stick as if it was a standard LB agar plate
- L.B.O. stick was incubated overnight at 37C
- 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
- June 29, 2010
- Followed procedure from June 26, 2010 to put new LB agar into the L.B.O. tube
- Approximately 1cm was sliced off the top of the new LB agar with an ethanol-sterilized razor blade
- L.B.O. stick was incubated overnight at 37C
Observations:
- no growth observed, tube must have been sterile
- June 30, 2010
- Streaked green colony from a plate of Cambridge parts
- L.B.O. stick was incubated overnight at 37C
Observations:
- green streak and individual green colonies observed, along with a distinct E.coli smell
- July 5, 2010
- Approximately 1cm was sliced off the top of the used LB agar with an ethanol-sterilized razor blade
- Streaked red, RFP-containing colony onto the LB agar surface.
- 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
Jello and Gelatine Plates
Used two types of Jello:
- Jell-o brand, raspberry flavor:
- sweetened with aspartame (no sugar added)
- 125mg sodium per 2.5g
- 1g protein per 2.5g
- 2. Knox brand Gelatine
Procedure (July 26, 2010): Jello:
- Stir two cups boiling water into 10g powder until dissolved
- pour into petri dishes
- chill for 2 hours until set
Gelatine:
- stir 175ml milli-Q water into 15ml gelatine
- stir in 175ml hot milli-Q and pour into petri dishes
- put in cold room until set (approx 2hours)
After 2 hours (Gelatine and Jello):
- Spread BBa_k274003(dark green stain of E.coli) onto 2 jello and 2 gelatine plates
- incubate overnight at 37C
Results: Both Jello and Gelatine plates had liquefied and no growth was seen.
