Team:Hawaii/Ligation of pRL1383a Parts

From 2008.igem.org

(Difference between revisions)
(New page: {{Team:Hawaii/Header}} ==Ligation of Parts== * The BioBrick parts of pRL1383a are to be ligated in a series of experiments. == Methods == #<strong> Round 1 of Restriction Digest and ...)
(adding experimetns from 8/11)
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{| border="1"
{| border="1"
-
|+ '''Restriction Digest & Ligation: <strong>Round 1</strong>'''
+
|+ '''Restriction Digest'''
!width="100"|Name
!width="100"|Name
-
!width="100"|Digest 1
+
!width="100"|size
-
!width="100"|Digest 2
+
!width="100"|enzyme
-
!width="100"|Ligation to
+
!width="100"|quantity
-
!width="100"|Product
+
!width="100"|date of digest
-
|-
+
-
|mob
+
-
|XbaI & PstI
+
-
|n/a
+
-
|[http://partsregistry.org/Part:BBa_B0024 BBa_B0024]
+
-
|TT-mob
+
|-
|-
|rep
|rep
 +
|3.3kb
|XbaI & PstI
|XbaI & PstI
-
|n/a
+
|5ng/ul
-
| [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]
+
|8/11/08
-
|P<sub>lac</sub>-rep
+
|-
|-
|oriV
|oriV
 +
|415bp
|XbaI & PstI
|XbaI & PstI
-
|n/a
+
|5ng/ul
-
|[http://partsregistry.org/Part:pSB1A3 pSB1A3]
+
|8/11/08
-
|pSB1A3::oriV
+
|-
|-
-
|''aadA'' region
+
|''aadA'' (pRL1383a)
 +
|806bp
|XbaI & PstI
|XbaI & PstI
-
|n/a
+
|5ng/ul
-
|[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]
+
|8/11/08
-
|P<sub>lac</sub>-aadA
+
|-
|-
-
|oriT
+
|''aadA'' (BB)
 +
|806bp
|XbaI & PstI
|XbaI & PstI
-
|n/a
+
|5ng/ul
-
|[http://partsregistry.org/Part:pSB1A3 pSB1A3]
+
|8/11/08
-
|pSB1A3::oriT
+
|-
|-
-
|pSB1A3
+
|oriT
 +
|~125bp
|XbaI & PstI
|XbaI & PstI
|n/a
|n/a
-
|see oriT and oriV
+
|8/27/08
-
|see oriT and oriV
+
|-
|-
-
|[http://partsregistry.org/Part:BBa_B0024 BBa_B0024]
+
|P1 lytic Region
-
|SpeI
+
|1.3kb
-
|PstI
+
|EcoRI & SpeI
-
|see mob region above
+
|5ng/ul
-
|see mob region above
+
|8/11/08
|-
|-
-
|[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]  
+
|[http://partsregistry.org/Part:BBa_B0030 BBa_B0030]
-
|SpeI
+
|2094bp
-
|PstI
+
|SpeI & PstI
-
|see rep and aadA region above
+
|4ng/ul
-
|see rep and aadA region above
+
|8/11/08
 +
|-
 +
|[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
 +
|3318bp
 +
|EcoRI & XbaI
 +
|5.7ng/ul
 +
|8/11/08
 +
|-
 +
|[http://partsregistry.org/Part:pSB1A2 pSB1A2]
 +
|2094bp
 +
|SpeI & PstI
 +
|5.5ng/ul
 +
|8/11/08
 +
|-
 +
|[http://partsregistry.org/Part:BBa_I14032 BBa_I14032]
 +
|4462bp
 +
|
 +
|n/a
 +
|n/a
|}
|}
-
#In gel purification of ligation products
+
#<strong>Ligation </strong>
-
 
+
-
#<strong> Round 2 of Restriction Digest and Ligation </strong>
+
{| border="1"
{| border="1"
-
|+ '''Restriction Digest & Ligation: <strong>Round 2</strong>'''
+
|+ ''' Ligation'''
!width="100"|Name
!width="100"|Name
-
!width="100"|Digest 1
 
-
!width="100"|Digest 2
 
-
!width="100"|Ligation to
 
-
!width="100"|Product
 
|-
|-
-
|TT-mob
+
|rep(80ng)+B0030(25ng)
-
|EcoRI
+
-
|SpeI
+
-
| [http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
+
-
|TT-mob-TT
+
|-
|-
-
|P<sub>lac</sub>-aadA
+
|oriV(20ng)+pSB1A2(50ng)
-
|EcoRI
+
-
|SpeI
+
-
| [http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
+
-
|P<sub>lac</sub>-aadA -TT
+
|-
|-
-
|[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
+
|aadA (BB)
-
|EcoRI
+
-
|XbaI
+
-
|see mob and aadA above
+
-
|see mob and aadA above
+
|}
|}

Revision as of 01:02, 14 August 2008

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

Contents

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
Name size enzyme quantity date of digest
rep 3.3kb XbaI & PstI 5ng/ul 8/11/08
oriV 415bp XbaI & PstI 5ng/ul 8/11/08
aadA (pRL1383a) 806bp XbaI & PstI 5ng/ul 8/11/08
aadA (BB) 806bp XbaI & PstI 5ng/ul 8/11/08
oriT ~125bp XbaI & PstI n/a 8/27/08
P1 lytic Region 1.3kb EcoRI & SpeI 5ng/ul 8/11/08
[http://partsregistry.org/Part:BBa_B0030 BBa_B0030] 2094bp SpeI & PstI 4ng/ul 8/11/08
[http://partsregistry.org/Part:BBa_B0015 BBa_B0015] 3318bp EcoRI & XbaI 5.7ng/ul 8/11/08
[http://partsregistry.org/Part:pSB1A2 pSB1A2] 2094bp SpeI & PstI 5.5ng/ul 8/11/08
[http://partsregistry.org/Part:BBa_I14032 BBa_I14032] 4462bp n/a n/a
  1. Ligation
Ligation
Name
rep(80ng)+B0030(25ng)
oriV(20ng)+pSB1A2(50ng)
aadA (BB)
  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]

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