Team:University of Sheffield /Wet Lab

From 2008.igem.org

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==Plasmids and Primers==
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{| align="center" border="1"
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! colspan="5"|Primers
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|-
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|BarA homology
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sequences on
 +
pKD13 Kanamycin
 +
resistant cassette
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|Forward:
 +
 +
5' - TGA TGA TTC TGA TCC TGG CAC CGA CCG TCC TTA TTG ATT CCG GGG ATC CGT CGA CC - 3' 
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 +
|TM (50mM NaCL):  71.8 °C 
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|GC Content:  55.4% 
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|Molecular Weight (Calculated):  17149.1
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|-
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|
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|Reverse:
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5' - CGT TGA CTT CGG GCG TCA CGA CGC GAG AGG AAA TAC GTG TAG GCT GGA GCT GCT TC - 3' 
 +
 
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|TM (50mM NaCL):  72.6 °C 
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|GC Content:  58.9% 
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|Molecular Weight (Calculated):  17377.2
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|-
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|RFP insertion
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into PGA operon
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= PGA homolgy on
 +
RFP cassette
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|Forward:
 +
 +
5' - TAA TTA TAC TCA CCA GCA TCA GGA GAT ATT TAT TTC CAT TAC GTA ACA TAT TTA TCC TTA TTA TTA AGC TAC TAA AGC GT - 3' 
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 +
|TM (50mM NaCL):  65.6 °C 
 +
|GC Content:  28.8% 
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|Molecular Weight (Calculated):  24492
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|-
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|
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|Reverse:
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5' - AAC TGG CGC GGT TTT GCT GGA TTC GGT TAT GCC GAT GGA CAA TTT AGC GAA GGA AAA GGG ATG GCT TCC TCC GAA GAC GT - 3' 
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 +
|TM (50mM NaCL):  73.3 °C 
 +
|GC Content:  51.2% 
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|Molecular Weight (Calculated):  24870.1
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|-
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|
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Revision as of 00:00, 29 October 2008

UniShefBanner.jpg


Introduction Our project Modelling Wet Lab Our team Timetable Miscellaneous


Introduction Protocols


Wet Lab

Wet lab sessions were where the theory now needed to come into practise. As mainly first years however, there was training to be done before we could get started.

Click here, or on the navigation bar, for the overall timetable of our wet lab sessions.

Click here, or on the sub-navigation bar, for almost all the protocols we used in our wet lab sessions.

The lab books below contain the more detailed jobs carried out by each member of the team.

Lab Books

Dimtry's Lab Book

Eva's Lab Book

Gosia's Lab Book

Hammad's Lab Book

Rosie's Lab Book

Sam's Lab Book


Plasmids and Primers

Primers
BarA homology

sequences on pKD13 Kanamycin resistant cassette

Forward:

5' - TGA TGA TTC TGA TCC TGG CAC CGA CCG TCC TTA TTG ATT CCG GGG ATC CGT CGA CC - 3'

TM (50mM NaCL): 71.8 °C GC Content: 55.4% Molecular Weight (Calculated): 17149.1
Reverse:

5' - CGT TGA CTT CGG GCG TCA CGA CGC GAG AGG AAA TAC GTG TAG GCT GGA GCT GCT TC - 3'

TM (50mM NaCL): 72.6 °C GC Content: 58.9% Molecular Weight (Calculated): 17377.2
RFP insertion

into PGA operon = PGA homolgy on RFP cassette

Forward:

5' - TAA TTA TAC TCA CCA GCA TCA GGA GAT ATT TAT TTC CAT TAC GTA ACA TAT TTA TCC TTA TTA TTA AGC TAC TAA AGC GT - 3'

TM (50mM NaCL): 65.6 °C GC Content: 28.8% Molecular Weight (Calculated): 24492
Reverse:

5' - AAC TGG CGC GGT TTT GCT GGA TTC GGT TAT GCC GAT GGA CAA TTT AGC GAA GGA AAA GGG ATG GCT TCC TCC GAA GAC GT - 3'

TM (50mM NaCL): 73.3 °C GC Content: 51.2% Molecular Weight (Calculated): 24870.1






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