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From 2010.igem.org

Lab work

Alkane Degradation

PCR Amplification

K398007A, K398008A, K398009A, K398010A, K398017A, K398018A, K398019A, B0015 were amplified with the universal primers G00100 and G00101. The product was put on 1% agarose gel:

Lane Description:

#	Description	Expected Length (bp)	Primers	Status	Remarks
M1	SmartLadder	n/a	n/a	n/a
1	PCR product of 007	1616	G00101 + G00101	✔
2	PCR product of 008	473	G00101 + G00101	✔
3	PCR product of 009	482	G00101 + G00101	✔
4	PCR product of 010	1505	G00101 + G00101	✔
5	PCR product of 017	1625	G00101 + G00101	✔
6	PCR product of 018	1052	G00101 + G00101	✔
7	PCR product of 019	1796	G00101 + G00101	✔
8	PCR product of B0015	445	G00101 + G00101	✔

The PCR products were subsequently digested:

#	Sample	Enzyme 1	Enzyme 2	Buffer	BSA	Needed fragment
1	40 μL PCR product 007	EcoRI	SpeI	2 (BioLabs)	✓	‘E–007–S’
2	40 μL PCR product 008	XbaI	PstI	2 (BioLabs)	✓	‘X–008-P’
3	40 μL PCR product 009	EcoRI	SpeI	2 (BioLabs)	✓	‘E–009–S’
4	40 μL PCR product 010	XbaI	PstI	2 (BioLabs)	✓	‘X–010-P’
5	40 μL PCR product 017	EcoRI	SpeI	2 (BioLabs)	✓	‘E–009–S’
6	40 μL PCR product 018	XbaI	PstI	2 (BioLabs)	✓	‘X–018-P’
7	40 μL PCR product 019	EcoRI	SpeI	2 (BioLabs)	✓	‘E–019-S’
8	40 μL PCR product B0015	XbaI	PstI	2 (BioLabs)	✓	‘X–B0015-P’