Team:Hawaii/Construction of pRL1383a

From 2008.igem.org

(Difference between revisions)
(→Parts)
Line 10: Line 10:
|+ '''Units of pRL1383a Construct'''
|+ '''Units of pRL1383a Construct'''
!width="100"|Name
!width="100"|Name
-
!width="100"|Status
+
!width="300"|Status
-
!width="100"|Quantity
+
!width="300"|Quantity
-
!width="100"|Individual construct
+
!width="300"|Individual construct
|-
|-
|OriV
|OriV

Revision as of 06:45, 31 July 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Contents

Construction of pRL1383a

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

Parts

Units of pRL1383a Construct
Name Status Quantity Individual construct
OriV PCR amplified 7/30 ? pSB1A3::OriV
rep region PCR amplified 7/29 ~960ng Plac-RBS-rep
omega interposon PCR amplified 7/30 ? pSB1A3::OriV
aadA gene PCR amplified 7/29 128ng Plac-RBS-aadA-TT
oriT Verified in vector 7/30 0(need plasmid prep, glycerol stock) pSB1A3::OriT
Plac[http://partsregistry.org/Part:BBa_I14032 BBa_I14032] verified by colony PCR need glycerol stock and plasmid prep Plac-RBS-aadA-TT, Plac-RBS-rep
RBS, efficiency 1.0[http://partsregistry.org/Part:BBa_B0034 BBa_B0034] plasmid available ? need to check Plac-RBS-aadA-TT, Plac-RBS-rep
Terminator [http://partsregistry.org/Part:BBa_B0015 BBa_B0015] plasmid prep 7/30 needs quantification Plac-RBS-aadA-TT
BioBrick Base Vector[http://partsregistry.org/Part:BBa_I51020 BBa_I51020] plasmid prep 7/30 needs quantification all parts will be inserted

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.


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]

Retrieved from "http://2008.igem.org/Team:Hawaii/Construction_of_pRL1383a"