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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 I 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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