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| | #* [[Team:Imperial_College_London/Wetlab/Protocols/Restriction|In Vitro Restriction Assay]] | | #* [[Team:Imperial_College_London/Wetlab/Protocols/Restriction|In Vitro Restriction Assay]] |
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| - | {| style="color:#CCC; background-color:#325d97;" cellpadding="6" cellspacing="0" border="3"
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| - | ! Section
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| - | ! Assay
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| - | ! Overview and Aims
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| - | | <b>1.Calibration Curves</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/Abs| TOP10 Growth]]
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| - | | * To produce a calibration curve to aid in the normalising of absorbance values. The relation of absorbance reading to number of cells varies with different cell strains. We are therefore doing one for Top-10. <br>
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| - | | <b>1.Promoter Characterisation</b>
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| - | | Blah
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| - | | To find
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| - | | <b>2.Auto-Induction</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/Glucose delay|Glucose Time Delay]]
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| - | | * Characterise the tunable time duration it takes before GFP expression (M2 activation)
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| - | | <b>2.Auto-Induction</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/IPTG-RFP |IPTG ]]
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| - | * Characterise Lac promoter by varying concentrations of IPTG
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| - | * Determine the IPTG concentration that allows for maximal protein production while still being non-toxic to the cell
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| - | *The cells will be grown until OD= 0.7.
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| - | *The RFP value will be monitered, although it is the GFP values that are more critical in this assay.
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| - | *OD and fluorescence data for GFP which can be converted in [http://partsregistry.org/cgi/measurement/new_batch.cgi Specific Promoter Units (SPUs)].
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| - | *The secondary carbon source will be taken from the previous experiment (see Secondary carbon source selection for CRP promoter)
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| - | *The IPTG concentrations will be taken from the previous experiment (see Determining concentration of IPTG)
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| - | [[Team:Imperial_College_London/Wetlab/See Glucose time delay for details]]
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| - | | <b>3.Polypeptide Production</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/IPTGgrowth |IPTG Toxicity]]
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| - | | To investigate the effect of our IPTG inducer on growth of our cultures.
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| - | * Measure growth rates at 28 degrees Celsius on minimal growth media so that subsequent testing timings etc. can be streamlined
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| - | * Determine the effect of IPTG toxicity on growth w/o any protein production complications
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| - | |- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
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| - | | <b>3.Polypeptide Production</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/Cellulase |Cellulase]]
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| - | | Aims
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| - | |- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
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| - | | <b>3.Polypeptide Production</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/PAH |PAH]]
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| - | | Aims
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| - | |- style="color:#333; background-color:#CCCCFF;" cellpadding="6" cellspacing="0" border="1"
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| - | | <b>5.Colanic Acid Encapsulation</b>
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| - | | Colanic Acid
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| - | | Colanic acid biosynthesis is both time consuming and metabolically expensive. If the E.ncapsulator is to be used in an industrial setting, then colanic acid mediated protection must be highly efficient. Protective efficiency can be defined as the percentage increase in fluorescence per µl of colanic acid produced (when compared to control cells). The following assay can be used to elucidate this parameter.
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| - | | <b>6.Trehalose</b>
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| - | | Colanic Acid
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| - | | asddaf
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| - | | <b>7.Thermoinduction</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/Thermoinduction |Thermoinduction]]
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| - | | ghj
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| - | | <b>8.Genome Restriction</b>
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| - | | [[Team:Imperial_College_London/Wetlab/Protocols/Restriction |In Vitro Restriction]]
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| - | | * By running restriction digests on the genome of E.coli strains, we can investigate the efficiency of our restriction enzyme, taqI and dpnII, on genome deletion.
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| - | * Run parallel digests with our chassis of choice (TOP10) and a Dam negative control (of which the DNA should not be methylated) to investigate the effects of methylation on cleavage.
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| - | |}
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| | =Autoinduction assays= | | =Autoinduction assays= |
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