Team:TorontoMaRSDiscovery/Notebook

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=April 27, 2009=
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[[image:To_igem_wiki_banner.jpg|965px]]
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#Received from Rosa (SPiT):
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{| style="color:white;background-color:#99CCFF;" height:100px cellpadding="2" cellspacing="0" border="0" width="100%" align="center" class="menu"
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#*TM0785
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!align="center"|[[Team:TorontoMaRSDiscovery|Home]]
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#**Plasmid containing encapsulin
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!align="center"|[[Team:TorontoMaRSDiscovery/Team|The Team]]
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#**Recommend transfect into bacteria and re-sequence
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!align="center"|[[Team:TorontoMaRSDiscovery/Project|The Project]]
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#**See email note regarding sequence error
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!align="center"|[[Team:TorontoMaRSDiscovery/Parts|BioBricks]]
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#*0.5 microliters TMG DNA 100 microgram/microliter
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!align="center"|[[Team:TorontoMaRSDiscovery/Modeling|Modelling]]
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#**Use 0.4 microliter for 50 microliter PCR reaction
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!align="center"|[[Team:TorontoMaRSDiscovery/Bioinformatics|Bioinformatics]]
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#**This is thumotoga maritime genomic DNA for purpose of re-cloning <Br />
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!align="center"|[[Team:TorontoMaRSDiscovery/Safety|Safety]]
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#Microcentrifuge tubes 1 and 2 placed in -20 freezer
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!align="center"|[[Team:TorontoMaRSDiscovery/Notebook|Notebook]]
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|}
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<br>
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=May 15, 2009=
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==Monthly Notebook==
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#pH buffers received from VWR Mississauga
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<ul>
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#*pH 4 buffer (red) 500 ml
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/April April]
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#*pH 7 buffer (yellow) 500 ml
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/May May]
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#*pH 10 buffer (blue) 500 ml<Br />''Above are used for pH/mV Meter calibration''
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/June June]
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#pH/mV meter calibrated according to manual – recorded in index
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/July July]
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#Ethanol solution (70%) made from 85% ethanol
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/Augst August]
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/September September]
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<li>[https://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook/October October]
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</ul>
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=2009-05-19=
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==Summary of Results==
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1. 2L of TE buffer made (10X TE)
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a. Recipe for 2L from stock solution (10X TE)
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i. 10 ml 1M Tris-HCl pH 7.5-80.0 x 2 = 20 ml
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ii. 2 ml 0.5 M EDTA pH 8.0 x 2 = 4 ml
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iii. 988 ml ddH20 x 2 = 1976 ml
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- To make Tris-HCl, Tris base is used and adjusted to desired pH using HCl
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{| border="1" cellpadding="5" cellspacing="0"
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- 1 M Tris base is required of which 20 ml is required – thereby 2.4228 g Tris base (due to scale limitations, 2.42g of Tris Base was used
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|'''Part Type'''
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|'''Arbitrary Name'''
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2. 500 ml of 1 M Tris Base made
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|'''Registry Code'''
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- mass of Tris used = 1M/1000 ml x 500 ml x 121.14 g/mol = 60.57 g
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|'''Construct Used For'''
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- volume of water used = 500 ml
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|'''Status'''
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|'''Transformants Stocked?'''
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3. 250 ml of 0.5 M EDTA solution was made
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|'''Antibiotic Resistant, Backbone Plasmid Visualized?'''
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- mass of EDTA used = 36.53 g
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|'''Part Visualized?'''
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- observations: EDTA did not dissolve in ddH2O on heat and being stirred
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|'''Sequenced?'''
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|----
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10 MINUTE BRIEFING (INSERT)
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|rowspan="2"|
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'''New Parts'''
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2009-05-21
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|Encapsulin (Enc)
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|K192000
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- retrieved autoclaved ddH20, glycerol solution
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|Encapsulin
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|Confirmed
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Gel Electrophoresis (test run)  
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|Yes
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|Yes
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- 1% gel: 0.5 g agar mixed with 50 ml of 1X TBE
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|Yes
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- 10X TBE diluted with 5 ml TBE + 45 ml ddH20 – did not use all 50 ml for 1 gel (gel will be too thick – make 2 instead)
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|Yes
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- Loading Dye: add glycerol solution to 25 mg bromophenol blue then add dH20 to 10 ml (to make 6X)  
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|----
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- Running gel: match wells to black side, run at 120 mA
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|eCFPtgt
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|K192001
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Visualize Gel in UV
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|CFP target protein
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1. Turn power on
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|Synthesized by Mr Gene
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2. Gel in machine face up
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|No
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3. Close door securely
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|No
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4. Turn white light on
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|No
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5. Adjust zoom, contrast, focus from black dial on top of machine
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|Yes
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6. Turn white light off (turns on UV)
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|----
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7. Press ‘live’ toggle – acq. Should be 0.4 sec.
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|rowspan="13"|
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8. Print if desired or save on floppy disk
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'''Registry Parts'''
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9. Turn power off
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|1
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10. Dispose of gel in proper container
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|JJ23100
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11. Close door
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|Control, Encapsulin
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|Confirmed
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2009-05-25
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|Yes
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|Yes
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- made overnight bacteria culture (200 ml LB: 20 microliters DB3.1)  
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|Too small
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|No
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2009-05-26
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|----
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|2
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|B0034
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|Control, Encapsulin
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|Confirmed
 +
|Yes
 +
|Yes
 +
|Too small
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|No
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|----
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|3
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|C0040
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|Control
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|Confirmed
 +
|Yes
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|Yes
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|Yes
 +
|No
 +
|----
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|4
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|C0012
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|Control
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|Confirmed
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|Yes
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|Yes
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|Yes
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|No
 +
|----
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|5
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|B0015
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|Control, Encapsulin
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|Confirmed
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|Yes
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|Yes
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|Yes
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|No
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|----
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|C (Amp + C)
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|pSB1AC3
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|Assembly
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|Confirmed
 +
|Yes
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|Yes
 +
|Yes
 +
|No
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|----
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|C (C only)
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|pSB1C3
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|Assembly
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|Confirmed
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|Yes
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|Yes
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|Yes
 +
|No
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|----
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|K (ccdb)
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|pSB1AK3
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|Assembly
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|Confirmed
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|Yes
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|Yes
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|Yes
 +
|No
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|----
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|K (RFP)
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|pSB1K3
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|Assembly
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|Confirmed
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|Yes
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|Yes
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|Yes
 +
|No
 +
|----
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|Tet (ccdb)
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|pSB1AT3
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|Assembly
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|Confirmed
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|Yes
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|Yes
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|Yes
 +
|No
 +
|----
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|Tet (RFP)
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|pSB1T3
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|Assembly
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|Confirmed
 +
|Yes
 +
|Yes
 +
|Yes
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|No
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|----
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|7
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|J13002
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|Encapsulin
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|Transfected
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|Yes
 +
|No
 +
|No
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|No
 +
|----
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|Amp
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|pSB1A3
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|Assembly
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|Transfected
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|Yes
 +
|No
 +
|No
 +
|No
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|-
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|rowspan="7"|
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'''Assembled Parts'''
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|----
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|Enc in C (Amp+C)
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|
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|Submission, Encapsulin
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|Confirmed
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|Yes
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|Yes
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|Yes
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|Yes
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|----
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|1+2 in C (Amp+C)
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|
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|Control, Encapsulin
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|Confirmed
 +
|Yes
 +
|Yes
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|Too small
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|No
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|----
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|3+2 in C (Amp+C)
 +
|
 +
|Control
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|Confirmed
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|Yes
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|Yes
 +
|Yes
 +
|No
 +
|----
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|4+5 in Tet (ccdb)
 +
|
 +
|Control
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|Confirmed
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|Yes
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|Yes
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|Yes
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|No
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|----
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|1+2+3+2 in K (RFP)
 +
|
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|Control
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|Transfected
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|Yes
 +
|No
 +
|No
 +
|No
 +
|----
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|1+2+Enc in K
 +
|
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|Encapsulin
 +
|Confirmed
 +
|Yes
 +
|Yes
 +
|Yes
 +
|In Progress
 +
|----
 +
|}

Latest revision as of 03:05, 22 October 2009

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Home The Team The Project BioBricks Modelling Bioinformatics Safety Notebook


Monthly Notebook

Summary of Results

Part Type Arbitrary Name Registry Code Construct Used For Status Transformants Stocked? Antibiotic Resistant, Backbone Plasmid Visualized? Part Visualized? Sequenced?

New Parts

Encapsulin (Enc) K192000 Encapsulin Confirmed Yes Yes Yes Yes
eCFPtgt K192001 CFP target protein Synthesized by Mr Gene No No No Yes

Registry Parts

1 JJ23100 Control, Encapsulin Confirmed Yes Yes Too small No
2 B0034 Control, Encapsulin Confirmed Yes Yes Too small No
3 C0040 Control Confirmed Yes Yes Yes No
4 C0012 Control Confirmed Yes Yes Yes No
5 B0015 Control, Encapsulin Confirmed Yes Yes Yes No
C (Amp + C) pSB1AC3 Assembly Confirmed Yes Yes Yes No
C (C only) pSB1C3 Assembly Confirmed Yes Yes Yes No
K (ccdb) pSB1AK3 Assembly Confirmed Yes Yes Yes No
K (RFP) pSB1K3 Assembly Confirmed Yes Yes Yes No
Tet (ccdb) pSB1AT3 Assembly Confirmed Yes Yes Yes No
Tet (RFP) pSB1T3 Assembly Confirmed Yes Yes Yes No
7 J13002 Encapsulin Transfected Yes No No No
Amp pSB1A3 Assembly Transfected Yes No No No

Assembled Parts

Enc in C (Amp+C) Submission, Encapsulin Confirmed Yes Yes Yes Yes
1+2 in C (Amp+C) Control, Encapsulin Confirmed Yes Yes Too small No
3+2 in C (Amp+C) Control Confirmed Yes Yes Yes No
4+5 in Tet (ccdb) Control Confirmed Yes Yes Yes No
1+2+3+2 in K (RFP) Control Transfected Yes No No No
1+2+Enc in K Encapsulin Confirmed Yes Yes Yes In Progress
Retrieved from "http://2009.igem.org/Team:TorontoMaRSDiscovery/Notebook"