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Revision as of 01:05, 4 September 2012 (view source) Sarah.grixti (Talk | contribs) (→Media Prep) ← Older edit		Revision as of 01:07, 4 September 2012 (view source) Sarah.grixti (Talk | contribs) (→Media Prep) Newer edit →
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	===[[Team:Macquarie_Australia/Protocols/ MediaPrep | Media Prep]]===		===[[Team:Macquarie_Australia/Protocols/ MediaPrep | Media Prep]]===
-	*'''LB media: (1L total)	+	*'''Further Media Preparation'''
-	Ingredients: Tryptone 10g, Yeast extract 5g, NaCl 10g, MilliQ water to 1000ml.	+
-		+
-	Methods: Dissolved 10g tryptone, 5g yeast extract and 10g NaCl in 800 mL MilliQ water, making use of a magnetic stirrer. Once dissolved, brought volume up to 1L using MilliQ water. Autoclaved 1000ml of the solution (121°C, 15 min, standard liquid cycle).'''	+

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Revision as of 01:07, 4 September 2012

Contents 1 Tuesday 7/8/12 1.1 Making liquid media, plates & buffers 1.2 Designing Gibson Assembly Fragments 2 Tuesday 14/8/12 2.1 Making Competent Cells 2.2 Outreach Planning Labwork 3 Wednesday 15/8/12 4 Tuesday 21/8/12 4.1 Outreach Planning Labwork 4.2 Making Plates 5 Tuesday 28/8/12 5.1 Outreach Planning 5.2 Outreach Lab Work 5.3 Competent Cell Testing 6 Friday 31/8/12 6.1 gBlock fragments 7 Tuesday 04/09/12 7.1 Gibson Assembly 7.2 School Visit 7.3 Competent Cell Protocol 7.4 Media Prep

Tuesday 7/8/12

(Click on headings to visit methods)

Making liquid media, plates & buffers

• Liquid LB Media

• SOC Solution

• SOB Solution

• LB Agar Plates

Ampicillin LB Agar Plates: 31 plates

Chloramphenicol LB Agar Plates: 33 Plates

Kanamyacin LB Agar Plates: 32 Plates

• TB buffer.

• TAE buffer.

• EDTA buffer.

Designing Gibson Assembly Fragments

• Haemoxygenase Gene
• Deinococcus radiodurans Bacteriophytochrome Gene
• Agrobacterium tumefaciens Bacteriophytochrome Gene

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Tuesday 14/8/12

(Click on headings to visit methods)

Making Competent Cells

• Choosing Biobricks
• Making competent cells
• Transformations

Outreach Planning Labwork

• Open Day

• Secondary Education Seminars

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Wednesday 15/8/12

Our open day preparation did not go as planned. Most of our transformations were unsuccessful, we believe this was because SOC was added after the heat shock. As disappointing as this was, there was one little piece of good news.

A single colony of Top 10 E.coli containing GFP insert was identified on a plate of LB agar media.

LB agar and nutrient agar plates were struck using this colony. The colony was also used to inoculate 5mL of nutrient broth. Plates and stock were then incubated at 37°C overnight. Both of the streaked plates and the broth showed significant growth of E.coli colonies which could fluoresce when exposed to UV light.

Unfortunately, Qantas declined to sponsor the effort of getting our team to Hong Kong. This was soon forgotten when the Macquarie Faculty of Medicine, Agilent and the Defence Science Institute (thanks Yagiz) both agreed to sponsor our team. Hong Kong here we come.

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Tuesday 21/8/12

Outreach Planning Labwork

• Secondary education seminars
• Reattempting glowing bacteria

Making Plates

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Tuesday 28/8/12

Outreach Planning

• High School Presentations: Preparation for 4th September
• Open Day: Preparation for 8th September at Macquarie University
• Writing the Abstract for the project due 7th September

Outreach Lab Work

• Fluorescent Bacteria

Competent Cell Testing

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Friday 31/8/12

gBlock fragments

• gBlock fragments from IDT

Tuesday 04/09/12

Gibson Assembly

• Gibson Assembly Protocol

School Visit

• Outreach

Competent Cell Protocol

• Generation of Competent Cells

Media Prep

• Further Media Preparation