Sensor: Week 8 July 5-8
From 2011.igem.org
(Difference between revisions)
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==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" | ||
Line 13: | Line 26: | ||
|- | |- | ||
|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 | + | ==Wednesday, July 6== |
- | + | ===cI Repressor Test Assembly, Day 28=== | |
+ | The sensor group attempted to place two ribosome binding sites behind the cI repressor. | ||
{|border="1" | {|border="1" | ||
- | ! | + | !Protocol |
- | + | |colspan="2" align="center"|'''Part Involved with Protocol''' | |
|- | |- | ||
- | | | + | |Insert PCR |
- | |align="center"| | + | |colspan="2" align="center"|B0033<br />B0032 |
|- | |- | ||
- | | | + | |rowspan="2"|Restriction Digest |
- | |align="center"| | + | |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. | ||
+ | |} | ||
[[Team:Penn_State/Notebook| Back to Notebook]] | [[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. |