Sensor: Week 11 July 25-29

From 2011.igem.org

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(Created page with "==Monday, July 25== {|border="1" !Protocol |align="center" colspan="2"|'''Part Involved with Protocol''' |- |rowspan="2"|Restriction Digest |Insert using EcoRI and SpeI: |B0034 |...")
 
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|Transformation/Plating
|Transformation/Plating
|colspan="2"|The above ligation was transformed into<br />Escherichia coli cells and plated onto<br />kanamycin resistant plates.
|colspan="2"|The above ligation was transformed into<br />Escherichia coli cells and plated onto<br />kanamycin resistant plates.
 +
|}
 +
O<sub>R</sub>+K64800 and O<sub>R</sub> were both grown up in culture to be used as freezer stocks.
 +
==Tuesday, July 26==
 +
{|border="2"
 +
!Protocol
 +
|align="center" colspan="2"|'''Part Involved with Protocol'''
 +
|-
 +
|rowspan="2"|Restriction Digest
 +
|Inserts using EcoRI and SpeI:
 +
|B0031 + C0051 (R)<br />B0030 + C0051 (R)<br />B0034 + C0051 (R)
 +
|-
 +
|Vector using EcoRI and XbaI:
 +
|O<sub>R</sub>+K648000
 +
|-
 +
|Ligation
 +
|colspan="2"|B0030 + C0051 + Or + K648000<br />B0031 + C0051 + Or + K648000<br />B0034 + C0051 + Or + K648000
 +
|-
 +
|Transformation/Plating
 +
|colspan="2"|The above ligations were transformed into Escherichia<br />coli cells and plated onto kanamycin resistant plates.
 +
|}
 +
The B0034+tev protease assembly from Monday, July 25 was verified on an agarose gel. The gel showed a band that corresponded to the correct size, so the colonies used for the gel were grown up in culture. The sequencing results came back positive.
 +
==Wednesday, July 27==
 +
{|border="2"
 +
!Protocol
 +
|align="center" colspan="2"|'''Part Involved with Protocol'''
 +
|-
 +
|Insert PCR
 +
|colspan="2"|O<sub>R</sub>
 +
|-
 +
|rowspan="2"|Restriction Digest
 +
|Insert using EcoRI and SpeI:
 +
|O<sub>R</sub>
 +
|-
 +
|Vector using EcoRI and XbaI:
 +
|34Cro
 +
|-
 +
|Ligation
 +
|colspan="2"|O<sub>R</sub>+34Cro
 +
|-
 +
|Transformation/Plating
 +
|colspan="2"|The above ligation was transformed into<br />Escherichia coli cells and plated onto<br />kanamycin resistant plates.
 +
|}
 +
The following constructs (assembled Tuesday, July 26) were verified on an agarose gel:
 +
{|
 +
|B0030 + C0051 + Or + K648000
 +
|-
 +
|B0031 + C0051 + Or + K648000
 +
|-
 +
|B0034 + C0051 + Or + K648000
 +
|}
 +
 +
''Results:'' The gel indicated that the plasmids were the correct size. The colonies used for the gel were grown up in culture.
 +
 +
==Thursday, July 28==
 +
The constructs assembled on Tuesday, July 26, yielded poor sequencing results. Thus, the assemblies will have to be repeated.
 +
 +
The O<sub>R</sub>+34Cro construct assembled Wednesday, July 27, was run on an agarose gel. The gel showed that the assembly failed and will have to be repeated.
 +
 +
{|border="1"
 +
!Protocol
 +
|align="center" colspan="2"|'''Part Involved with Protocol'''
 +
|-
 +
|Insert PCR
 +
|colspan="2|B0015
 +
|-
 +
|rowspan="2"|Restriction Digest
 +
|Insert using XbaI and PstI:
 +
|B0015
 +
|-
 +
|Vector using SpeI and PstI:
 +
|B0034+tev protease
 +
|-
 +
|Ligation
 +
|colspan="2"|B0034+tev protease+B0015
 +
|-
 +
|Transformation/Plating
 +
|colspan="2"|The above ligation was transformed into<br />Escherichia coli cells and plated onto<br />kanamycin resistant plates.
 +
|}
 +
 +
The 34Cro part was grown up in culture to be used as freezer stock.
 +
 +
==Friday, July 29==
 +
The following plasmids were extracted using the Omega Bio-Tek miniprep kit and sent to the sequencing center:
 +
{|
 +
KOr  + Cro (A)<br />KOr + Cro (B)
 +
|}
 +
The following parts were amplified through PCR:
 +
{|
 +
C0051 + B0030<br />C0051 + B0031<br />C0051 + B0034
 +
|}
[[Team:Penn_State/Notebook| Back to Notebook]]
[[Team:Penn_State/Notebook| Back to Notebook]]

Latest revision as of 19:52, 4 August 2011

Contents

Monday, July 25

Protocol Part Involved with Protocol
Restriction Digest Insert using EcoRI and SpeI: B0034
Vector using EcoRI and XbaI: tev protease
Ligation B0034+tev protease
Transformation/Plating The above ligation was transformed into
Escherichia coli cells and plated onto
kanamycin resistant plates.

OR+K64800 and OR were both grown up in culture to be used as freezer stocks.

Tuesday, July 26

Protocol Part Involved with Protocol
Restriction Digest Inserts using EcoRI and SpeI: B0031 + C0051 (R)
B0030 + C0051 (R)
B0034 + C0051 (R)
Vector using EcoRI and XbaI: OR+K648000
Ligation B0030 + C0051 + Or + K648000
B0031 + C0051 + Or + K648000
B0034 + C0051 + Or + K648000
Transformation/Plating The above ligations were transformed into Escherichia
coli cells and plated onto kanamycin resistant plates.

The B0034+tev protease assembly from Monday, July 25 was verified on an agarose gel. The gel showed a band that corresponded to the correct size, so the colonies used for the gel were grown up in culture. The sequencing results came back positive.

Wednesday, July 27

Protocol Part Involved with Protocol
Insert PCR OR
Restriction Digest Insert using EcoRI and SpeI: OR
Vector using EcoRI and XbaI: 34Cro
Ligation OR+34Cro
Transformation/Plating The above ligation was transformed into
Escherichia coli cells and plated onto
kanamycin resistant plates.

The following constructs (assembled Tuesday, July 26) were verified on an agarose gel:

B0030 + C0051 + Or + K648000
B0031 + C0051 + Or + K648000
B0034 + C0051 + Or + K648000

Results: The gel indicated that the plasmids were the correct size. The colonies used for the gel were grown up in culture.

Thursday, July 28

The constructs assembled on Tuesday, July 26, yielded poor sequencing results. Thus, the assemblies will have to be repeated.

The OR+34Cro construct assembled Wednesday, July 27, was run on an agarose gel. The gel showed that the assembly failed and will have to be repeated.

Protocol Part Involved with Protocol
Insert PCR B0015
Restriction Digest Insert using XbaI and PstI: B0015
Vector using SpeI and PstI: B0034+tev protease
Ligation B0034+tev protease+B0015
Transformation/Plating The above ligation was transformed into
Escherichia coli cells and plated onto
kanamycin resistant plates.

The 34Cro part was grown up in culture to be used as freezer stock.

Friday, July 29

The following plasmids were extracted using the Omega Bio-Tek miniprep kit and sent to the sequencing center:

KOr + Cro (A)
KOr + Cro (B)

The following parts were amplified through PCR:

C0051 + B0030
C0051 + B0031
C0051 + B0034

Back to Notebook

Retrieved from "http://2011.igem.org/Sensor:_Week_11_July_25-29"