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From 2009.igem.org

• Miniprep of fluid cultures

Nanodrop results:

Part	concentration (ng/μl)	260/280 λ
SamS I	53,9	1,87
SamS II	39,4	2,16
SamS III	57,1	2,07
EF I	76,8	1,94
EF II	83,4	2,04
EF III	61,5	2,01

• Restriction:

-We performed 2 simultaneous digests: one to verify the length of the product (Restriction2Test) using SAMS II and EF III and another to prepare for ligation (Restriction4Real) with promotor and terminator using SAMS III and EF II

-We cut with the 2 enzymes separately, allowing a one hour incubation in between. EF: E/S SAMS: E/S

-Restriction volumes:

Restriction2Test

Part	DNA µl	MQ µl
SamS II	12,7	17,3
EF III	8,1	21,9

Restriction4Real

Part	DNA µl	MQ µl
SamS III	8,7	21,3
EF II	6,0	24,0

• Terminator was also digested

Terminator: E/X

Part	DNA µl	MQ µl
Terminator	2,7	27,3

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