Sensor: Week 4 June 6-11

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Revision as of 15:59, 15 June 2011

Monday

cI Repressor Test Assembly, Day 12

The minipreps prepared on 6/4 (K648000+K648001, B0030+C0051, B0032+C0051) were sent to sequencing. The sequencing results showed that the K648000+K648001 and B0030+C0051 assemblies worked. However, the B0032+C0051 assembly only produced vector background. Thus, stock should be made of the K648000+K648001 and B0030+C0051 constructs and the B0032+C0051 assembly should be redone.

Cro and O_R Insert into C3 Vector, Day 1

The sensor group attempted to insert the Cro and O_R parts created through PCR on 6/3 into separate C3 vectors through amplified insert assembly.

Assembly Part	Part Involved with Insert Step
Restriction Digest	Inserts using XbaI and PstI:	O_R Cro
Restriction Digest	Vector using XbaI and PstI:	C3+violacein
Ligation	O_R+C3 Cro+C3
Transformation/Plating	The above ligations were transformed into Escherichia coli cells and plated onto Chloramphenicol resistant plates.

Tuesday

cI Repressor Test Assembly, Day 13

The sensor group worked to make stock of their correct assemblies. The following constructs were transformed into Escherichia coli cells and allowed to grow in culture in order to make freezer stock.

K648000+K648001

B0030+C0051

B0034+C0051

Freezer stock was made from the B0031+C0051 culture as well.

Cro and O_R Insert into C3 Vector, Day 2

All of the colonies on the plates were purple, meaning that the insert never inserted into the vector. Thus, Cro and O_R must be inserted into a different vector.

Wednesday

cI Repressor Test Assembly, Day 14

The B0034+C0051 construct was sent to sequencing. Freezer stocks were made from the cultures made 6/7 (listed above).

Cro and O_R Insert into K3 Vector, Day 1

Since the C3 vector did not accept the Cro and O_R inserts, the sensor group attempted to insert Cro and O_R into the K3 vector.

Assembly Step	Part Involved with Assembly Step
Restriction Digest	Inserts using XbaI and PstI:	O_R Cro
Restriction Digest	Vector using XbaI and PstI:	K3
Ligation	O_R+K3 Cro+K3
Transformation/Plating	The above ligations were transformed into Escherichia coli cells and plated onto Kanamycin resistant plates.

Back to Notebook

@@ Line 20: / Line 20: @@
 |-
 |Transformation/Plating
-|colspan="2"|The above ligations were transformed into<br />Escherichia coli cells and plated onto<br />chloramphenicol resistant plates.
+|colspan="2"|The above ligations were transformed into<br />Escherichia coli cells and plated onto<br />Chloramphenicol resistant plates.
 |}
@@ Line 30: / Line 30: @@
 <blockquote>B0034+C0051</blockquote>
 Freezer stock was made from the B0031+C0051 culture as well.
+===Cro and O<sub>R</sub> Insert into C3 Vector, Day 2===
+All of the colonies on the plates were purple, meaning that the insert never inserted into the vector. Thus, '''Cro and O<sub>R</sub> must be inserted into a different vector.'''
 ==Wednesday==
-===
+===cI Repressor Test Assembly, Day 14===
+The B0034+C0051 construct was sent to sequencing. Freezer stocks were made from the cultures made 6/7 (listed above).
+===Cro and O<sub>R</sub> Insert into K3 Vector, Day 1===
+Since the C3 vector did not accept the Cro and O<sub>R</sub> inserts, the sensor group attempted to insert Cro and O<sub>R</sub> into the K3 vector.
+{|border="1"
+!Assembly Step
+|colspan="2" align="Center"|'''Part Involved with Assembly Step'''
+|-
+|rowspan="2"|Restriction Digest
+|Inserts using XbaI and PstI:
+|O<sub>R</sub><br />Cro
+|-
+|Vector using XbaI and PstI:
+|K3
+|-
+|Ligation
+|colspan="2"|O<sub>R</sub>+K3<br />Cro+K3
+|-
+|Transformation/Plating
+|colspan="2"|The above ligations were transformed into<br />Escherichia coli cells and plated onto<br />Kanamycin resistant plates.
+|}