|
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| Line 6: |
Line 6: |
| | * [[Melbourne/Lab Notebook Weeks 13-16|Weeks 13-16]] | | * [[Melbourne/Lab Notebook Weeks 13-16|Weeks 13-16]] |
| | * [[Melbourne/Lab Notebook Weeks 17-18|Weeks 17-18]] | | * [[Melbourne/Lab Notebook Weeks 17-18|Weeks 17-18]] |
| - |
| |
| - | ==Week 1==
| |
| - | *25 June 2007: Prepared LB agar plates Amp & Kana.
| |
| - | *25 June 2007: [[Melbourne/IGEM 2007 kit|Resuspended the following from Registry plates]] & [[Melbourne/Transformation Protocol shorter|Transformed (shorter protocol)]] into competent DH5alpha cells.
| |
| - | *#[[Melbourne/BBa_I15008|'''P2 21A''']] - (BBa_I15008, ho1, Kan) -> No colonies on plates
| |
| - | *#[[Melbourne/BBa_I15009|'''P2 21C''']] - (BBa_I15009, PcyA, Kan) ->No colonies on plates
| |
| - | *#[[Melbourne/BBa_R0084|'''P1 11H''']] - (BBa_R0084, OmpR positive promoter, Amp)->Small number of colonies.
| |
| - |
| |
| - |
| |
| - |
| |
| - | *26 June 2007: [[Melbourne/IGEM 2007 kit|Resuspended the following from Registry plates]] & [[Melb:Transformation Protocol|Transformed]] into Joe's competent DH5alpha cells.
| |
| - | *#[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034, RBS, Amp)
| |
| - | *#[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051, c1 protein, Amp)
| |
| - | *#[[Melbourne/BBa_B0010|'''P2 3P''']] (BBa_B0010, Terminator, Amp)
| |
| - |
| |
| - | *26 June 2007: Streaked the following cells:
| |
| - | *#[[Melbourne/pJS010|'''pJS010''']] (from solid agar, Amp)
| |
| - | *#[[Melbourne/Fusion|'''Fusion protein''']] (from glycerol stock, Amp?)
| |
| - | *#[[Melbourne/BBa_I15010|'''BBa_I15010''']] (from solid agar, Kan)
| |
| - |
| |
| - | *27 June 2007: [[Melb:Transformation Protocol|Transformed]] into Joe's competent DH5alpha cells.
| |
| - | *#[[Melbourne/BBa_I15008|'''P2 21A''']] (Kan)
| |
| - | *#[[Melbourne/BBa_I15009|'''P2 21C''']] (Kan)
| |
| - |
| |
| - |
| |
| - | <font color="red"><b>Liquid culture</b></font>
| |
| - | #[[Melb:Growing up cells|Prepared]] 30mL of Amp LB (100ug/mL). Prepared 5mL Kan LB (50mg/mL).
| |
| - | #Aliquoted 5mL Amp LB into 6 50mL falcon tubes
| |
| - | #To the Amp LB aliquots single transformed colonies from plates of the following were introduced:
| |
| - | #*[[Melbourne/pJS010|'''pJS010''']]
| |
| - | #*[[Melbourne/Fusion|'''Fusion''']]
| |
| - | #*[[Melbourne/BBa_B0034|'''P1 3O''']]
| |
| - | #*[[Melbourne/BBa_C0051|'''P1 5G''']]
| |
| - | #*[[Melbourne/BBa_B0010|'''P2 3P''']]
| |
| - | #*[[Melbourne/BBa_R0084|'''P1 11H''']]
| |
| - | #To the Kan LB a single colony from the transformation plate of [[Melbourne/BBa_I15010|'''BBa_I15010''']] was introduced.
| |
| - | #Cells incubated at 37degrees with shaking overnight.
| |
| - |
| |
| - | <font size=3><b>28 June 2007
| |
| - | </b>
| |
| - | </font>
| |
| - |
| |
| - | <font color="red"><b>Miniprep</b></font>
| |
| - | #[[Melb:Miniprep protocol|Minipreped]] the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
| |
| - | #*[[Melbourne/pJS010|'''pJS010''']]
| |
| - | #*[[Melbourne/Fusion|'''Fusion''']]
| |
| - | #*[[Melbourne/BBa_B0034|'''P1 3O''']]
| |
| - | #*[[Melbourne/BBa_C0051|'''P1 5G''']]
| |
| - | #*[[Melbourne/BBa_B0010|'''P2 3P''']]
| |
| - | #*[[Melbourne/BBa_R0084|'''P1 11H''']]
| |
| - | #*[[Melbourne/BBa_I15010|'''I15010''']]
| |
| - | #Stored in -20 freezer
| |
| - |
| |
| - |
| |
| - | <font color="red"><b>Liquid culture</b></font>
| |
| - | #[[Melb:Growing up cells|Cultured]] the following cells from transformed plates:
| |
| - | #*[[Melbourne/BBa_R0084|'''P1 11H''']] (BBa_R0084)
| |
| - | #*[[Melbourne/BBa_I15010|'''BBa_I15010''']]
| |
| - | #*[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034)
| |
| - | #*[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051)
| |
| - | #*[[Melbourne/BBa_I15008|'''P1 21A''']] (BBa_I15008)
| |
| - | #*[[Melbourne/BBa_I15009|'''P1 21C''']] (BBa_I15009)
| |
| - |
| |
| - | <font size=3><b>29 June 2007
| |
| - | </b>
| |
| - | </font><br>
| |
| - | <font color="red"><b>Miniprep</b></font>
| |
| - | #[[Melb:Miniprep protocol|Minipreped]] the cultures grown overnight after putting aside 1mL(sterile) for glycerol stocks:
| |
| - | #*[[Melbourne/BBa_R0084|'''P1 11H''']] (BBa_R0084)
| |
| - | #*[[Melbourne/BBa_I15010|'''BBa_I15010''']]
| |
| - | #*[[Melbourne/BBa_B0034|'''P1 3O''']] (BBa_B0034)
| |
| - | #*[[Melbourne/BBa_C0051|'''P1 5G''']] (BBa_C0051)
| |
| - | #*[[Melbourne/BBa_I15008|'''P1 21A''']] (BBa_I15008)
| |
| - | #*[[Melbourne/BBa_I15009|'''P1 21C''']] (BBa_I15009)
| |
| - |
| |
| - | #Stored in -20 freezer
| |
| - |
| |
| - | ==Week 2==
| |
| - | *2 July 2007: [[Melbourne/IGEM 2007 kit|Resuspended the following from Registry plates]] & [[Melb:Transformation Protocol|Transformed]] into Joe's competent DH5alpha cells.
| |
| - | *#Q04510
| |
| - | *#E0241
| |
| - | *#E0040
| |
| - | *#B0014
| |
| - | *#J61035
| |
| - |
| |
| - | *3 July 2007: [[Melb:Transformation Protocol|Re Transformed]] into Joe's competent DH5alpha cells. Each Plate had additional 40uL of 50mg/ml Ampicillan spread on surface.
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J''']] -> Three colonies -> grew in liquid culture 4 july
| |
| - | *#[[Melbourne/BBa_E0040|'''P1 5H''']] -> Multiple colonies at edge -> did not grow in overnight liquid culture 4 july 'poor amp spreading'
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L''']] -> Three colonies -> grew in liquid culture 4 july
| |
| - |
| |
| - | *4 July 2007:
| |
| - |
| |
| - | <font color="red"><b>Ampicillin Plates</b></font>
| |
| - | *Made up 25 Ampicillin plates using 400mL LB agar prepared on the 25th of June.
| |
| - | **Used 100mg/ml Ampicillin stock from Gooley Lab - In bag in left freezer near our lab bench.
| |
| - |
| |
| - | <font color="red"><b>Transformation</b></font> <BR>
| |
| - | [[Melbourne/Transformation Protocol|Transformed]] the following and grew on new ampicillin plates
| |
| - | *[[Melbourne/BBa_E0040|'''P1 5H''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L''']]
| |
| - | **Also Plated an untransformed control (subjected to transformation protocol in absense of DNA) and a blank plate control (no cells)
| |
| - |
| |
| - | <font color="red"><b>Liquid Culture</b></font><BR>
| |
| - | *[[Melbourne/Growing up cells|Cultured]] 2 colonies from each of the following transformed plates and labelled as follows
| |
| - | **[[Melbourne/BBa_B0010|'''P2 3P 1''']]
| |
| - | **[[Melbourne/BBa_B0010|'''P2 3P 2''']]
| |
| - | **[[Melbourne/BBa_I15010|'''I15010 1''']]
| |
| - | **[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | **[[Melbourne/pJS010|'''pJS010 1''']]
| |
| - | **[[Melbourne/pJS010|'''pJS010 2''']]
| |
| - | **[[Melbourne/Fusion|'''Fusion 1''']]
| |
| - | **[[Melbourne/Fusion|'''Fusion 2''']]
| |
| - | **[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | **[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | **[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | **[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | **[[Melbourne/BBa_Q04510|'''P2 13K 1''']]
| |
| - | **[[Melbourne/BBa_Q04510|'''P2 13K 2''']]
| |
| - | **[[Melbourne/BBa_B0014|'''P1 1G 1''']]
| |
| - | **[[Melbourne/BBa_B0014|'''P1 1G 2''']]
| |
| - | **[[Melbourne/BBa_R0084|'''P1 11H 1''']]
| |
| - | **[[Melbourne/BBa_R0084|'''P1 11H 2''']]
| |
| - | **[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | **[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | **suspected contaminating colony from [[Melbourne/BBa_J61035|'''P4 8J''']]
| |
| - |
| |
| - | <font size=3><b>5 July 2007
| |
| - | </b>
| |
| - | </font>
| |
| - |
| |
| - | <font color="red"><b>Miniprep</b></font><BR>
| |
| - | *1mL culture from the 4th of Julyput aside for glycerol stocks under sterile conditions. Labelled with todays date 5/7
| |
| - | *miniprepped the following overnight cultures set up on the 4th of July. Final elution was performed with TE buffer prepared on 25/06/07 rather than nuclease free water. Labelled with todays date 5/7.
| |
| - | **[[Melbourne/BBa_B0010|'''P2 3P 1''']]
| |
| - | **[[Melbourne/BBa_B0010|'''P2 3P 2''']]
| |
| - | **[[Melbourne/BBa_I15010|'''I15010 1''']]
| |
| - | **[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | **[[Melbourne/pJS010|'''pJS010 1''']]
| |
| - | **[[Melbourne/pJS010|'''pJS010 2''']]
| |
| - | **[[Melbourne/Fusion|'''Fusion 1''']]
| |
| - | **[[Melbourne/Fusion|'''Fusion 2''']]
| |
| - | **[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | **[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | **[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | **[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | **[[Melbourne/BBa_Q04510|'''P2 13K 1''']](eluted in 130uL due to accidental double application of 50uL elution)
| |
| - | **[[Melbourne/BBa_Q04510|'''P2 13K 2''']]
| |
| - | **[[Melbourne/BBa_B0014|'''P1 1G 1''']]
| |
| - | **[[Melbourne/BBa_B0014|'''P1 1G 2''']]
| |
| - | *the following liquid cultures were not miniprepped due to failure (no growth)
| |
| - | **[[Melbourne/BBa_R0084|'''P1 11H 1''']]
| |
| - | **[[Melbourne/BBa_R0084|'''P1 11H 2''']]
| |
| - | **[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | **[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | **suspected contaminating colony from [[Melbourne/BBa_J61035|'''P4 8J''']]
| |
| - | <BR>
| |
| - | <font color="red"><b>Digest</b></font> <BR>
| |
| - | Performed the following [[Melbourne/Diagnostic Digest|digests]] on DNA from the above miniprep <BR><BR>
| |
| - | <font color="green"><b>EcoR1/Pst1 with buffer 3</b></font>
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 1''']]
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | *[[Melbourne/BBa_Q04510|'''P2 13K 1''']]
| |
| - | *[[Melbourne/BBa_Q04510|'''P2 13K 2''']]
| |
| - | <font color="green"><b>EcoR1/HaeII in buffer 2</b></font>
| |
| - | *[[Melbourne/BBa_B0010|'''P2 3P 1''']]
| |
| - | *[[Melbourne/BBa_B0010|'''P2 3P 2''']]
| |
| - | *[[Melbourne/BBa_B0014|'''P1 1G 1''']]
| |
| - | *[[Melbourne/BBa_B0014|'''P1 1G 2''']]
| |
| - | <font color="green"><b>XbaI/SpeI in buffer 2</b></font>
| |
| - | *[[Melbourne/pJS010|'''pJS010 1''']]
| |
| - | *[[Melbourne/pJS010|'''pJS010 2''']]
| |
| - |
| |
| - | Incubated for 2hours at 37 degrees, before addition of loading dye and storage at -20
| |
| - |
| |
| - | <font color="red"><b>Transformation</b></font><BR>
| |
| - | *[[Melbourne/Transformation Protocol|Transformed]] P1 11H from resuspended DNA.
| |
| - |
| |
| - | <font color="red"><b>Liquid Culture</b></font>
| |
| - | *[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | *[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - |
| |
| - | <font size=3><b>6 July 2007
| |
| - | </b>
| |
| - | </font>
| |
| - |
| |
| - | <font color="red"><b>Digest Gel</b></font><BR>
| |
| - | * Prepared 20 lane 100mL [[Melbourne/Preparing an agarose gel|agarose gel]] with 0.5xTBE buffer.
| |
| - | *[[Melbourne/Loading a DNA gel|Loaded]] 20uL of digest samples in the following lane order
| |
| - | *#1kb+ ladder
| |
| - | *#[[Melbourne/BBa_I15010|'''I15010 1''']]
| |
| - | *#[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | *#[[Melbourne/BBa_Q04510|'''P2 13K 1''']]
| |
| - | *#[[Melbourne/BBa_Q04510|'''P2 13K 2''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *#[[Melbourne/BBa_B0014|'''P1 1G 1''']]
| |
| - | *#[[Melbourne/BBa_B0014|'''P1 1G 2''']]
| |
| - | *#[[Melbourne/BBa_B0010|'''P2 3P 1''']]
| |
| - | *#[[Melbourne/BBa_B0010|'''P2 3P 2''']]
| |
| - | *#[[Melbourne/pJS010|'''pJS010 1''']]
| |
| - | *#[[Melbourne/pJS010|'''pJS010 2''']]
| |
| - | *Ran for 1.5hours at 95V
| |
| - |
| |
| - | <font color="red"><b>Miniprep</b></font><BR>
| |
| - | *Put aside 1mL of liquid cultures set up on the 5th for glycerol stocks and labelled with todays date 6/7
| |
| - | *Miniprepped the remains of the cultures and labelled with todays date 6/7. Samples were eluted with TE buffer.
| |
| - | *[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | *[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - |
| |
| - |
| |
| - | <font color="red"><b>Digest</b></font><BR>
| |
| - | *[[Melbourne/Diagnostic Digest|Digested]] 5uL of each of the above miniprep DNA with EcoRI and PstI in buffer 3.
| |
| - | *Incubated for 2hours 25min at 37degrees
| |
| - | *Added 5uL 6x loading dye and stored at -20
| |
| - |
| |
| - | <font color="red"><b>Glycerol Stocks</b></font><BR>
| |
| - | The following [[Melbourne/Glycerol Stocks|glycerol stocks]] were made:
| |
| - | *Put aside from cultures 5/7 (labelled with this date)
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *#[[Melbourne/BBa_B0014|'''P1 1G 1''']]
| |
| - | *#[[Melbourne/BBa_B0014|'''P1 1G 2''']]
| |
| - | *Put aside from cultures 6/7 (labelled with this date)
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *#[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | *#[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | Stored at -80
| |
| - |
| |
| - | <font color="red"><b>Liquid Culture</b></font><BR>
| |
| - | [[Melbourne/Growing up cells|Cultured]] the following in 5mL LB
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 1''']] (Kan)
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | *[[Melbourne/BBa_R0084|'''P1 11H 1''']] (Amp)
| |
| - | *[[Melbourne/BBa_R0084|'''P1 11H 2''']]
| |
| - |
| |
| - | Also placed Kan plate in the incubator to test antibiotic efficiency - no growth on 8/7
| |
| - |
| |
| - | <font size=3><b>7 July 2007
| |
| - | </b>
| |
| - | </font>
| |
| - |
| |
| - | *Made 10x TAE buffer
| |
| - |
| |
| - | <font color="red"><b>Digest Gel</b></font><BR>
| |
| - | *Prepared 8 lane 60mL [[Melbourne/Preparing an agarose gel|agarose gel]] with 1xTAE buffer.
| |
| - | *[[Melbourne/Loading a DNA gel|Loaded]] 20uL of digest samples from 6/7 in the following lane order.
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 1''']]
| |
| - | *#[[Melbourne/BBa_J61035|'''P4 8J 2''']]
| |
| - | *#[[Melbourne/BBa_E0040|'''P1 5H 1''']]
| |
| - | *#[[Melbourne/BBa_E0040|'''P1 5H 2''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 1''']]
| |
| - | *#[[Melbourne/BBa_E0241|'''P2 15L 2''']]
| |
| - |
| |
| - | <font color="red"><b>Glycerol Stocks</b></font><BR>
| |
| - | The following [[Melbourne/Glycerol Stocks|glycerol stocks]] were made and dated 7/7:
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 1''']]
| |
| - | *[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | *[[Melbourne/BBa_R0084|'''P1 11H 1''']]
| |
| - | *[[Melbourne/BBa_R0084|'''P1 11H 2''']]
| |
| - | Stored at -80
| |
| - |
| |
| - | <font color="red"><b>Miniprep</b></font><BR>
| |
| - | *Miniprepped the remains of the cultures and labelled with todays date 7/7. Samples were eluted with TE buffer.
| |
| - |
| |
| - | <font color="red"><b>Digest</b></font><BR>
| |
| - | *[[Melbourne/Diagnostic Digest|Digested]] 5uL of each of the above miniprep DNA
| |
| - | **EcoRI/PstI in buffer 3
| |
| - | ***[[Melbourne/BBa_I15010|'''I15010 1''']] (Kan)
| |
| - | ***[[Melbourne/BBa_I15010|'''I15010 2''']]
| |
| - | ***[[Melbourne/BBa_R0084|'''P1 11H 1''']] (Amp)
| |
| - | ***[[Melbourne/BBa_R0084|'''P1 11H 2''']]
| |
| - | *Incubated for 3hours at 37degrees
| |
| - | *Added 5uL 6x loading dye and stored at -20
| |
| - |
| |
| - | <font size=3><b>8 July 2007
| |
| - | </b>
| |
| - | </font>
| |
| - |
| |
| - | <font color="red"><b>Transformation</b></font><BR>
| |
| - | Resuspended and [[Melbourne/Transformation Protocol|transformed]] the following
| |
| - | *[[Melbourne/BBa_R0082|'''P1 15P''']](BBa_R0082, Omp R+, Amp)
| |
| - | *[[Melbourne/BBa_R0083|'''P1 17H''']](BBa_R0083, truncated BBa_R0082 Omp R+, Amp)
| |
| - | *[[Melbourne/BBa_E0430|'''P1 11A''']](BBa_E0430, EYFP(RBS+,LVA-,term) Amp)
| |
| - | *[[Melbourne/BBa_E0840|'''P1 16E''']](BBa_E0430; RBS,GFP,term; Amp)
| |
| - | The following was also retransformed due to colonies on previous plate appearing to be contaminants and failure of plasmid isolation from these colonies.
| |
| - | *[[Melbourne/BBa_Q04510|'''P2 13K''']] (BBa_Q04510, c1 inverter, Kan)
| |
| - |
| |
| - | ==Week 3==
| |
| - | <font size=3><b>9 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Digested I15010(E/P),P1-11H(E/H),P2-15L 1.5hrs run
| |
| - | *liquid culture P1-11H,I15010,P1-15P,P1-16E,P1-17H,P2-13K
| |
| - | <font size=3><b>10 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Miniprep
| |
| - | *Digest P1-16E,P1-11A,P2-13K,I15010,P1-15P,P1-11H,P1-17H 1.0hrs run
| |
| - | *liquid culture I15010, P2
| |
| - | <font size=3><b>11 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Digest for ligation P1-15P(1)10/7,P1-11H 10/7,P1-17H(1)10/7,P1-16E(2)10/7,P1-11A(1)10/7
| |
| - | *loaded order X/P P1-11A,X/P P1-16E,S/P P1-15P,S/P P1-11H
| |
| - | ** Spe1(6uL)/Pst1(7.5uL)/AP(1.5uL), Buffer2 9uL, BSA 9uL, milliQ 27uL into 20uL aliquots with 10uL DNA.
| |
| - | ** Xbal1(3uL)/PstI(4uL),Buffer2 6uL,10XBSA 6uL,milliQ 21uL into 20uL aliquots with 10uL DNA
| |
| - | *Excise bands of interest and purify invitrogen
| |
| - | *liquid culture P1-11A,P1-15P 10ml
| |
| - | *Transform P2-21B,P2-23N,P3-20I into DB3.1 heat shock.
| |
| - | *Glycerol stocks P1-11A,P1-16E,P1-11H,P1-15P,P1-17H
| |
| - | <font size=3><b>12 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Ran Gel P1-11A,P1-16E,P1-15P,P1-11H
| |
| - | *miniprepped P1-11A,P1-15P
| |
| - | *Digest
| |
| - | *Ligate control=(2uL ligase buffer,1uL ligase,5uL vector(P1-15P),12uL H20)
| |
| - | *Ligate (2uL ligase buffer,1uL ligase,5uL vector(P1-15P),10uL insert(P1-11A),2uL H20)
| |
| - | *Liquid culture transformants 11/7
| |
| - | <font size=3><b>13 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *miniprep cultures from transformants 11/7
| |
| - | *Digestion P1-15P and P1-11A from 12/7/07 37degC 3hours 15 minutes stopped 5uL of 6X loading dye.
| |
| - | *Excise bands 800bp from P1-11A, 2Kbp from P1-15P and purified.
| |
| - | *Glycerol stocks of P3-20I,P2-21B,P2-23N
| |
| - | *Transform DH5a with ligation product
| |
| - | <font size=3><b>14 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Transform using 10uL of ligation reaction
| |
| - |
| |
| - | ==Week 4==
| |
| - | <font size=3><b>16 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *[[Melbourne/colony pcr|Colony PCR]]
| |
| - | <font size=3><b>18 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | *Purchased XbaI,EcoRI,PstI
| |
| - | *Miniprepped cultures 1,3,6 from 16/7
| |
| - | *Digestion with E/P
| |
| - | *Ran Gel:std,ctrl(from PCR reaction 1 earlier),Digest1,3,6,(P2-15P)ages ago,(P1-11A)ages ago?
| |
| - |
| |
| - | ==Week 5==
| |
| - | <font size=3><b>23 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | <font size=3><b>24 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | <font size=3><b>25 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | <font size=3><b>26 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | <font size=3><b>27 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | ==Week 6==
| |
| - | <font size=3><b>30 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>31 July 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>1 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>2 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>3 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>4 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | ==Week 7==
| |
| - | <font size=3><b>5 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>6 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>7 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>8 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>9 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>10 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>11 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | ==Week 8==
| |
| - | <font size=3><b>12 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>13 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>14 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>15 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>16 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>17 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>18 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | ==Week 9==
| |
| - | <font size=3><b>19 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>20 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>21 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>22 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>23 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>24 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>25 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - | ==Week 10==
| |
| - | <font size=3><b>26 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>27 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>28 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>29 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>30 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>31 Aug 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>1 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | ==Week 11==
| |
| - | <font size=3><b>2 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>3 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>4 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>5 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>6 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>7 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>8 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | ==Week 12==
| |
| - | <font size=3><b>9 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>10 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>11 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>12 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>13 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>14 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>15 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | ==Week 13==
| |
| - | <font size=3><b>16 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>17 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>18 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>19 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>20 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>21 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>22 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | ==Week 14==
| |
| - | <font size=3><b>23 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>24 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>25 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>26 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - | *Miniprepped the following
| |
| - | **TetR-RBS-P2,21A-ter A,B,C and D
| |
| - | **TetR-RBS-ComP-ter A,B,C and D
| |
| - | **GenRBS-ComA-ter A,B,C and D
| |
| - | **cI-L3 A,B,C and D
| |
| - |
| |
| - | <font size=3><b>27 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>28 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>29 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - |
| |
| - | ==Week 15==
| |
| - | <font size=3><b>30 Sept 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>1 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>2 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>3 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>4 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>5 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>6 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | ==Week 16==
| |
| - | <font size=3><b>7 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>8 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>9 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>10 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>11 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>12 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>13 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | ==Week 17==
| |
| - | <font size=3><b>14 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>15 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>16 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>17 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>18 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>19 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>20 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | ==Week 18==
| |
| - | <font size=3><b>21 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>22 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>23 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>24 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>25 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
| - |
| |
| - |
| |
| - | <font size=3><b>26 Oct 2007
| |
| - | </b>
| |
| - | </font><BR>
| |
