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	==June 2010==		==June 2010==
	===June 1/2010===		===June 1/2010===
-	JV quantified the amount of DNA in gels run to date using ImageJ software. Results to be posted in [[Team:Lethbridge/Notebook/Working_Plasmids|working plasmids box]].<br><br>	+	JV quantified the amount of DNA in gels run to date using ImageJ software. Results to be posted in [[Team:Lethbridge/Notebook/Working_Plasmids|working plasmids box]].<br>
		+	<b>Objective:</b> Transform plasmids into DH5&alpha;<br>
		+	<b>Method:</b> Follow [[Team:Lethbridge/Notebook/Protocols|competent cell transformation]] protocol to transform the following:<br>
		+	From our ligations:<br>
		+	*pLacI-sRBS-Lumazine-dT<br>
		+	*pLacI-sRBS-Lumazine-dT<br>
		+	*mms6 (A6)<br>
		+	*mms6 (B6)<br>
		+	*xylE (C4)<br>
		+	*xylE (B4)<br>
		+	From the 2010 Parts Distribution:<br>
		+	*ECFP (Bba_E0020)<br>
		+	*EYFP (Bba_E0030)<br>
		+	*BglII Endonuclease (Bba_K112106)<br>
	===June 2/2010===		===June 2/2010===

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Revision as of 00:14, 11 June 2010

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Contents 1 June 2010 1.1 June 1/2010 1.2 June 2/2010 1.3 June 3/2010

June 2010

June 1/2010

JV quantified the amount of DNA in gels run to date using ImageJ software. Results to be posted in working plasmids box.

Objective: Transform plasmids into DH5α
Method: Follow competent cell transformation protocol to transform the following:
From our ligations:

• pLacI-sRBS-Lumazine-dT
  
• pLacI-sRBS-Lumazine-dT
  
• mms6 (A6)
  
• mms6 (B6)
  
• xylE (C4)
  
• xylE (B4)
  

From the 2010 Parts Distribution:

• ECFP (Bba_E0020)
  
• EYFP (Bba_E0030)
  
• BglII Endonuclease (Bba_K112106)
  

June 2/2010

(In Lab: JV)

Objective: Isolate plasmid DNA (BBa_J33204) from DH5α cells and confirm results.

Method: "Mini-prep" the plasmid DNA using boiling lysis miniprep. Then restrict the DNA once and run on a 1% agarose gel (TAE).

Restriction Reaction

Ingredient	Volume(µL)
MilliQ H20 Water	15.75
Orange Buffer (10x)	2
pDNA (rbs-xylE)	2
EcoRI	0.25

Unrestricted Control

Ingredient	Volume(µL)
MilliQ H20 Water	16
Orange Buffer (10x)	2
pDNA (rbs-xylE)	2

I started ran the reaction for 80 minutes at 37^oC.

June 3/2010

Objective: Ligate pLacI to sRBS-Lum-dT using the three antibiotic assembly method (according to Tom Knight's protocol). Previous ligation had miniscule quantities of DNA in ligation, not surprising it didn't work.