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Revision as of 13:01, 23 July 2012 (view source) Elbrich (Talk | contribs) (Created page with "{{HeaderGroningen2012}}") ← Older edit		Revision as of 11:50, 25 July 2012 (view source) Elbrich (Talk | contribs) Newer edit →
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		+
		+
		+	Plan for today:
		+
		+	1. plasmid isolation from E.coli Dh5A
		+
		+	2. ligate alsT promoter and GFP into pSB1C3 and pSac-Cm (B. subtilis backbone)
		+
		+	3. E.coli transformation for both
		+
		+
		+	1. Plasmid isolation pSB1C3+insert from iGEM Groningen 2010
		+
		+	E.coli DH5a strong magenta = insert device OK!
		+
		+	plasmid concentration: 129,1 ng/ul
		+
		+
		+	Purification of PCR product using gel DNA extraction kit
		+
		+	diluted in 50ul TE (elution buffer)
		+
		+	concentration: 35,6 ng.ul
		+
		+
		+	Ligation pAlsT-GFP into backbone pSB1C3 (and pSac-Cm done next week 23/07)
		+
		+	Cut parts with restiction enzymes
		+
		+	alsT: EcoRI + SpeI
		+
		+	GFP: XbaI + PstI
		+
		+	backbone: EcoRI + PstI
		+
		+	Digestion reaction and subsequently ligation of:
		+	1) alsT150-GFP-BB
		+
		+	2) alsT150-GFP-BB
		+
		+	3) alsT150-GFP-BB
		+
		+	4) alsT150-GFP-BB

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Revision as of 11:50, 25 July 2012

Plan for today:

1. plasmid isolation from E.coli Dh5A

2. ligate alsT promoter and GFP into pSB1C3 and pSac-Cm (B. subtilis backbone)

3. E.coli transformation for both

1. Plasmid isolation pSB1C3+insert from iGEM Groningen 2010

E.coli DH5a strong magenta = insert device OK!

plasmid concentration: 129,1 ng/ul

Purification of PCR product using gel DNA extraction kit

diluted in 50ul TE (elution buffer)

concentration: 35,6 ng.ul

Ligation pAlsT-GFP into backbone pSB1C3 (and pSac-Cm done next week 23/07)

Cut parts with restiction enzymes

alsT: EcoRI + SpeI

GFP: XbaI + PstI

backbone: EcoRI + PstI

Digestion reaction and subsequently ligation of: 1) alsT150-GFP-BB

2) alsT150-GFP-BB

3) alsT150-GFP-BB

4) alsT150-GFP-BB