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From 2008.igem.org

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Contents 1 Ligation of Parts 2 Methods 3 Results 4 Discussion

Ligation of Parts

• The BioBrick parts of pRL1383a are to be ligated in a series of experiments.

Methods

1. Round 1 of Restriction Digest and Ligation

Restriction Digest & Ligation: Round 1

Name	Digest 1	Digest 2	Ligation to	Product
mob	XbaI & PstI	n/a	[http://partsregistry.org/Part:BBa_B0024 BBa_B0024]	TT-mob
rep	XbaI & PstI	n/a	[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]	Plac-rep
oriV	XbaI & PstI	n/a	[http://partsregistry.org/Part:pSB1A3 pSB1A3]	pSB1A3::oriV
aadA region	XbaI & PstI	n/a	[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]	Plac-aadA
oriT	XbaI & PstI	n/a	[http://partsregistry.org/Part:pSB1A3 pSB1A3]	pSB1A3::oriT
pSB1A3	XbaI & PstI	n/a	see oriT and oriV	see oriT and oriV
[http://partsregistry.org/Part:BBa_B0024 BBa_B0024]	SpeI	PstI	see mob region above	see mob region above
[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]	SpeI	PstI	see rep and aadA region above	see rep and aadA region above

1. In gel purification of ligation products

1. Round 2 of Restriction Digest and Ligation

Restriction Digest & Ligation: Round 2

Name	Digest 1	Digest 2	Ligation to	Product
TT-mob	EcoRI	SpeI	[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]	TT-mob-TT
Plac-aadA	EcoRI	SpeI	[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]	Plac-aadA -TT
[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]	EcoRI	XbaI	see mob and aadA above	see mob and aadA above

1. Transformation

1. Verification with Colony PCR

Results

• 
  

Discussion

• What was learned and how to do future experiments differently.

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