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From 2010.igem.org

Revision as of 10:00, 27 August 2010 (view source) Ptmvanboheemen (Talk | contribs) ← Older edit		Revision as of 08:45, 31 August 2010 (view source) Mvoges (Talk | contribs) Newer edit →
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		+	==Alkane degradation==
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		+	====Transformation====
		+	Yesterday's ligation mixes were transformed into TOP10 competent cells and plated out onto CAM plates.
		+
		+	====Ligation kinetics====
		+	Yesterday's ligation mixes were transformed into TOP10 competent cells and plated out onto CAM plates.
		+
		+	====Digestions====
		+
	==Alkane Degradation parallel attempt==		==Alkane Degradation parallel attempt==

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Revision as of 08:45, 31 August 2010

Contents 1 Strain Characterization 2 Alkane degradation 2.1 Transformation 2.2 Ligation kinetics 2.3 Digestions 3 Alkane Degradation parallel attempt

Strain Characterization

By Hugo

I'm working in some growth curves, today I tested a different amount of inoculum than yesterday.

Measurements!!!

Hour	Time	Glc	Glc+CC6	C8
11:43	0h	0.015	0.014	0.007
14:54	3h 11'	0.042	0.052	0.066
16:00	4h 17'	0.099	0.081	0.086
17:00	5h 17'	0.196	0.128	0.142
18:00	6h 17'	0.355	0.215	0.184
19:00	7h 17'	0.555	0.341	0.275
9:43	22h	1.473	1.366	1.195
11:43	24h	1.459	1.360	1.398

Alkane degradation

Transformation

Yesterday's ligation mixes were transformed into TOP10 competent cells and plated out onto CAM plates.

Ligation kinetics

Yesterday's ligation mixes were transformed into TOP10 competent cells and plated out onto CAM plates.

Digestions

Alkane Degradation parallel attempt

Yesterdays plates all contained colonies. But since this is a two way ligation, we expect a lot of religated backbones, so false positive colonies. That is why we performed a colony PCR.

We picked 5 colonies from each plate.

Lane description

#	Description	Expected size (bp)	OK?
1	Smartladder	n/a	Yes
2-6	014	3000+	No
7-11	020	2100+	No
12-16	304	342	Maybe
17-21	327	310	Maybe

Because the gels are inconclusive about biobrick 304 and 327 we decided to isolate the plasmids and send them to BaseClear for sequencing.