Glasgow/Wetlab/Gels

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!align=center|[https://2007.igem.org/Glasgow https://static.igem.org/mediawiki/2007/thumb/c/cc/Uog.jpg/50px-Uog.jpg] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#99CCFF size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]]
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!align=center|[[Image:Uog.jpg]] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]]
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|-
|-
|[[Glasgow/Wetlab/Week6#Friday 10th August 2007|10th Aug 2007 (2)]]
|[[Glasgow/Wetlab/Week6#Friday 10th August 2007|10th Aug 2007 (2)]]
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|Colony PCR after ligating (*m*), (*s*), DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 1.
+
|Colony PCR after ligating phzM, phzS, DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 1.
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|'''1.'''(*m*)<br>
+
|'''1.'''phzM<br>
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'''2.'''(*m*)<br>
+
'''2.'''phzM<br>
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'''3.'''(*s*)<br>
+
'''3.'''phzS<br>
-
'''4.'''(*s*)<br>
+
'''4.'''phzS<br>
-
'''5.'''(*s*)<br>
+
'''5.'''phzS<br>
-
'''6.'''(*s*)<br>
+
'''6.'''phzS<br>
'''7.'''neg control<br>
'''7.'''neg control<br>
'''8.'''neg control<br>
'''8.'''neg control<br>
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|-
|-
|[[Glasgow/Wetlab/Week6#Friday 10th August 2007|10th Aug 2007 (2)]]
|[[Glasgow/Wetlab/Week6#Friday 10th August 2007|10th Aug 2007 (2)]]
-
|Colony PCR after ligating (*m*), (*s*), DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 2.
+
|Colony PCR after ligating phzM, phzS, DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 2.
-
|'''1.'''(*s*)<br>
+
|'''1.'''phzS<br>
-
'''2.'''(*s*)<br>
+
'''2.'''phzS<br>
'''3.'''neg control<br>
'''3.'''neg control<br>
'''4.'''DntR<br>
'''4.'''DntR<br>
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|-
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|[[Glasgow/Wetlab/Week7#Monday 13th August 2007|13th Aug 2007 (4)]]
|[[Glasgow/Wetlab/Week7#Monday 13th August 2007|13th Aug 2007 (4)]]
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|PCR of genes (*a→g*), (*a→d*), and (*d→g*).
+
|PCR of genes phzA→phzG, phzA→phzD, and phzD→phzG. Wells in the gel were made larger by putting tape on the combs in order to yield more DNA in gel extraction.
-
|'''1A.'''(*a→g*) <br>
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|'''1A.'''phzA→phzG <br>
-
'''1B.'''(*a→g*) <br>
+
'''1B.'''phzA→phzG <br>
-
'''1C.'''(*a→g*) <br>
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'''1C.'''phzA→phzG <br>
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'''2A.'''(*a→g*) <br>
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'''2A.'''phzA→phzG <br>
-
'''2B.'''(*a→d*) <br>
+
'''2B.'''phzA→phzD <br>
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'''2C.'''(*d→g*) <br>
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'''2C.'''phzD→phzG <br>
|[[Image:2007-08-13 12hr 43min.JPG|220px]]
|[[Image:2007-08-13 12hr 43min.JPG|220px]]
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|-
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|[[Glasgow/Wetlab/Week7#Friday 17th August 2007|17th Aug 2007 (2-4)]]
|[[Glasgow/Wetlab/Week7#Friday 17th August 2007|17th Aug 2007 (2-4)]]
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|PstI digested minipreps of (*m*) and (*s*) in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 1.
+
|PstI digested minipreps of phzM and phzS in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 1.
-
|'''1.-3.'''(*m*)+4/6B<br>(PstI site mutated)<br>
+
|'''1.-3.'''phzM+4/6B<br>(PstI site mutated)<br>
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'''4.-6.'''(*m*)+3/20G<br>(PstI site mutated)<br>
+
'''4.-6.'''phzM+3/20G<br>(PstI site mutated)<br>
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'''7.-9.'''(*s*)+4/6B<br>(1st PstI site mutated)<br>
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'''7.-9.'''phzS+4/6B<br>(1st PstI site mutated)<br>
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'''10.-12.'''(*s*)+3/20G<br>(2nd PstI site mutated)<br>
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'''10.-12.'''phzS+3/20G<br>(2nd PstI site mutated)<br>
-
'''13.-15.'''(*s*)+4/6B<br>(2nd PstI site mutated)<br>
+
'''13.-15.'''phzS+4/6B<br>(2nd PstI site mutated)<br>
-
'''16.'''(*s*)+3/20G<br>(1st PstI site mutated)<br>
+
'''16.'''phzS+3/20G<br>(1st PstI site mutated)<br>
|[[Image:17thAug SDM1 PstI digest gel 1.JPG|220px]]
|[[Image:17thAug SDM1 PstI digest gel 1.JPG|220px]]
|-
|-
|[[Glasgow/Wetlab/Week7#Friday 17th August 2007|17th Aug 2007 (2-4)]]
|[[Glasgow/Wetlab/Week7#Friday 17th August 2007|17th Aug 2007 (2-4)]]
-
|PstI digested minipreps of (*m*) and (*s*) in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 2.
+
|PstI digested minipreps of phzM and phzS in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 2.
-
|'''1.-2.'''(*s*)+3/20G<br>(1st PstI site mutated)<br>
+
|'''1.-2.'''phzS+3/20G<br>(1st PstI site mutated)<br>
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'''3.'''(*m*)+4/6B<br>('''Before''' mutagenesis)<br>
+
'''3.'''phzM+4/6B<br>('''Before''' mutagenesis)<br>
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'''4.'''(*m*)+3/20G<br>('''Before''' mutagenesis)<br>
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'''4.'''phzM+3/20G<br>('''Before''' mutagenesis)<br>
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'''5.'''(*s*)+4/6B<br>('''Before''' mutagenesis)<br>
+
'''5.'''phzS+4/6B<br>('''Before''' mutagenesis)<br>
-
'''6.'''(*s*)+3/20G<br>('''Before''' mutagenesis)<br>
+
'''6.'''phzS+3/20G<br>('''Before''' mutagenesis)<br>
|[[Image:17thAug SDM1 PstI digest gel 2.JPG|220px]]
|[[Image:17thAug SDM1 PstI digest gel 2.JPG|220px]]
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|-
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|[[Glasgow/Wetlab/Week8#Thursday 23rd August 2007|23rd Aug 2007]]
|[[Glasgow/Wetlab/Week8#Thursday 23rd August 2007|23rd Aug 2007]]
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|(*s*) minipreps in 4/6B and 3/20G run after the second round of site-directed mutagenesis.
+
|phzS minipreps in 4/6B and 3/20G run after the second round of site-directed mutagenesis.
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|'''1.-5.'''(*s*)+4/6B<br>
+
|'''1.-5.'''phzS+4/6B<br>
-
'''6.-16.'''(*s*)+3/20G<br>
+
'''6.-16.'''phzS+3/20G<br>
|[[Image:Maija's Gel taken by Scott.JPG|200px]]
|[[Image:Maija's Gel taken by Scott.JPG|200px]]
|-
|-
|[[Glasgow/Wetlab/Week8#Thursday 23rd August 2007|23rd Aug 2007]]
|[[Glasgow/Wetlab/Week8#Thursday 23rd August 2007|23rd Aug 2007]]
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|PstI digested (*s*) minipreps in 4/6B and 3/20G vectors after the second round of site-directed mutagenesis.
+
|PstI digested phzS minipreps in 4/6B and 3/20G vectors after the second round of site-directed mutagenesis.
-
|'''1.-5.'''(*s*)+4/6B<br>
+
|'''1.-5.'''phzS+4/6B<br>
-
'''6.-16.'''(*s*)+3/20G<br>
+
'''6.-16.'''phzS+3/20G<br>
|[[Image:23rdAug 13hr 46min Maija.JPG|200px]]
|[[Image:23rdAug 13hr 46min Maija.JPG|200px]]
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{| valign=top cellpadding=3
{| valign=top cellpadding=3
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|-
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!align=center|[https://2007.igem.org/Glasgow https://static.igem.org/mediawiki/2007/thumb/c/cc/Uog.jpg/50px-Uog.jpg] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#99CCFF size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]]
+
!align=center|[[Image:Uog.jpg]] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]]
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|}

Latest revision as of 17:06, 22 October 2007

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Wetlab Log

Contents

Gels

Friday 10th August 2007

Date and Reference Description Key Gel
10th Aug 2007 (2) Colony PCR after ligating phzM, phzS, DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 1. 1.phzM

2.phzM
3.phzS
4.phzS
5.phzS
6.phzS
7.neg control
8.neg control
9.neg control

10thAug gel 1.JPG
10th Aug 2007 (2) Colony PCR after ligating phzM, phzS, DntR and Pu with the construction vectors 3/20G and 4/6B. Gel 2. 1.phzS

2.phzS
3.neg control
4.DntR
5.DntR
6.Pu
7.Pu
8.neg control
9.neg control
10.neg control
11.neg control

10thAug gel 2.JPG

Monday 13th August 2007

Date and Reference Description Key Gel
13th Aug 2007 (4) PCR of genes phzA→phzG, phzA→phzD, and phzD→phzG. Wells in the gel were made larger by putting tape on the combs in order to yield more DNA in gel extraction. 1A.phzA→phzG

1B.phzA→phzG
1C.phzA→phzG
2A.phzA→phzG
2B.phzA→phzD
2C.phzD→phzG

2007-08-13 12hr 43min.JPG

Friday 17th August 2007

Date and Reference Description Key Gel
17th Aug 2007 (2-4) PstI digested minipreps of phzM and phzS in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 1. 1.-3.phzM+4/6B
(PstI site mutated)

4.-6.phzM+3/20G
(PstI site mutated)
7.-9.phzS+4/6B
(1st PstI site mutated)
10.-12.phzS+3/20G
(2nd PstI site mutated)
13.-15.phzS+4/6B
(2nd PstI site mutated)
16.phzS+3/20G
(1st PstI site mutated)

17thAug SDM1 PstI digest gel 1.JPG
17th Aug 2007 (2-4) PstI digested minipreps of phzM and phzS in 4/6B and 3/20G vectors before and after the first round of site-directed mutagenesis. Gel 2. 1.-2.phzS+3/20G
(1st PstI site mutated)

3.phzM+4/6B
(Before mutagenesis)
4.phzM+3/20G
(Before mutagenesis)
5.phzS+4/6B
(Before mutagenesis)
6.phzS+3/20G
(Before mutagenesis)

17thAug SDM1 PstI digest gel 2.JPG

Thursday 23rd August 2007

Date and Reference Description Key Gel
23rd Aug 2007 phzS minipreps in 4/6B and 3/20G run after the second round of site-directed mutagenesis. 1.-5.phzS+4/6B

6.-16.phzS+3/20G

Maija's Gel taken by Scott.JPG
23rd Aug 2007 PstI digested phzS minipreps in 4/6B and 3/20G vectors after the second round of site-directed mutagenesis. 1.-5.phzS+4/6B

6.-16.phzS+3/20G

23rdAug 13hr 46min Maija.JPG
23rd Aug 2007 Colony PCR with "Pr_Prefix" and "XylR_Suffix" primers from Lynsey's transformations with Pr+XylR miniprep DNA. Gel 1. 1.-7.colonies from the plate Gel for Maija 1.JPG
23rd Aug 2007 Colony PCR with "Pr_Prefix" and "XylR_Suffix" primers from Lynsey's transformations with Pr+XylR miniprep DNA. Gel 2. 8.-12.colonies from the plate

13.neg control

Gel for Maija 2.JPG


Friday 24th August 2007

Date and Reference Description Key Gel
24th Aug 2007 (1) These are the miniprepped plasmids from Pr+XylR in TOPO vector. The seven colonies gave the right size product from colony PCR but failed to give right size bands after miniprepping. Wait for sequencing results. 1.-7.colonies from Pr+XylR in TOPO transformations 2007-08-24 12hr 54min Maija.JPG

Monday 27th August 2007

Date and Reference Description Key Gel
27th Aug 2007 (1) pDntA in 3/20G and 4/6B vectors digested with Roche's PvuI restriction enzyme. Results do not match up with expected fragment sizes, thus performing a PCR with "VF2" and "VR" primers. 1.pDntA+3/20G

2.pDntA+4/6B

2007-08-27 13hr 21min Maija.JPG
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