Sensor: Week 8 July 5-8

From 2011.igem.org

(Difference between revisions)
Line 1: Line 1:
==Tuesday, July 5==
==Tuesday, July 5==
 +
===cI Repressor Test Assembly, Day 27===
 +
''Sequencing Results:'' The following constructs were sent to sequencing to verify their DNA sequences:
 +
{|border="1"
 +
!Construct
 +
!Sequencing<br />Result
 +
|-
 +
|J23113 + B0034 + C0051
 +
|align="center"|Good
 +
|-
 +
|KOR + K648000
 +
|align="center"|Good
 +
|}
 +
===Clone O<sub>R</sub> and CRO===
===Clone O<sub>R</sub> and CRO===
{|border="1"
{|border="1"
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|-
|-
|Ligation
|Ligation
-
|colspan="2"|K3 + OR<br />K3 + CRO
+
|colspan="2" align="center"|K3 + OR<br />K3 + CRO
|-
|-
|Transformation/Plating
|Transformation/Plating
Line 19: Line 32:
|}
|}
-
===cI Repressor Test Assembly, Day 27===
+
==Wednesday, July 6==
-
''Sequencing Results:'' The following constructs were sent to sequencing to verify their DNA sequences:
+
===cI Repressor Test Assembly, Day 28===
 +
The sensor group attempted to place two ribosome binding sites behind the cI repressor.
{|border="1"
{|border="1"
-
!Construct
+
!Protocol
-
!Sequencing<br />Result
+
|colspan="2" align="center"|'''Part Involved with Protocol'''
|-
|-
-
|J23113 + B0034 + C0051
+
|Insert PCR
-
|align="center"|Good
+
|colspan="2" align="center"|B0033<br />B0032
|-
|-
-
|KOR + K648000
+
|rowspan="2"|Restriction Digest
-
|align="center"|Good
+
|Inserts using EcoRI and SpeI:
 +
|B0032<br />B0033
 +
|-
 +
|Vector using EcoRI and XbaI:
 +
|C0051
 +
|-
 +
|Ligation
 +
|colspan="2" align="center"|C0051 + B0032<br />C0051 + B0033
 +
|-
 +
|Transformation/Plating
 +
|colspan="2"|The above ligations were transformed into<br />Escherichia coli cells and plated onto<br />ampicillin resistant plates.
|}
|}
 +
===Clone O<sub>R</sub> and CRO, Take 2===
 +
No colonies grew on the kanamycin resistant plates from 7/5, so the cloning was attempted again.
-
 
+
{|border="1"
 +
!Protocol
 +
|colspan="2" align="center"|'''Part Involved with Protocol'''
 +
|-
 +
|rowspan="2"|Restriction Digest
 +
|Inserts using XbaI and PstI:
 +
|O<sub>R</sub><br />CRO
 +
|-
 +
|Vector using XbaI and PstI:
 +
|K3
 +
|-
 +
|Ligation
 +
|colspan="2" align="center"|K3 + OR<br />K3 + CRO
 +
|-
 +
|Transformation/Plating
 +
|colspan="2"|The above ligations were transformed into<br />Escherichia coli cells and plated onto<br />kanamycin resistant plates.
 +
|}
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[[Team:Penn_State/Notebook| Back to Notebook]]

Revision as of 18:03, 13 July 2011

Contents

Tuesday, July 5

cI Repressor Test Assembly, Day 27

Sequencing Results: The following constructs were sent to sequencing to verify their DNA sequences:

Construct Sequencing
Result
J23113 + B0034 + C0051 Good
KOR + K648000 Good

Clone OR and CRO

Protocol Part Involved with Protocol
Restriction Digest Inserts using XbaI and PstI: OR
CRO
Vector using XbaI and PstI: K3
Ligation K3 + OR
K3 + CRO
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto
kanamycin resistant plates.

Wednesday, July 6

cI Repressor Test Assembly, Day 28

The sensor group attempted to place two ribosome binding sites behind the cI repressor.

Protocol Part Involved with Protocol
Insert PCR B0033
B0032
Restriction Digest Inserts using EcoRI and SpeI: B0032
B0033
Vector using EcoRI and XbaI: C0051
Ligation C0051 + B0032
C0051 + B0033
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto
ampicillin resistant plates.

Clone OR and CRO, Take 2

No colonies grew on the kanamycin resistant plates from 7/5, so the cloning was attempted again.

Protocol Part Involved with Protocol
Restriction Digest Inserts using XbaI and PstI: OR
CRO
Vector using XbaI and PstI: K3
Ligation K3 + OR
K3 + CRO
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto
kanamycin resistant plates.


Back to Notebook

Retrieved from "http://2011.igem.org/Sensor:_Week_8_July_5-8"