Team:Queens Canada/Notebook/Week17

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Notebook - Week 17 and onwards

DateProtocolPeopleDNA (if relevant)Quantities and Parameters (if relevant)Notes on Protocol
19/08/2012PCR overlap extensionAndrewxylE XS and PP eCFP XS into nFF GFP + J04500  
19/08/2012DigestionAndrewSmtA and Fmt with XS  
20/08/2012GelDavidDP fmt XS, DP SmtA XS, eCFP full construct PCR product  
20/08/2012PCR overlap extensionAndrew   
23/08/2012Liquid culturesKevinC. thermo x-dockerin II pet21b, CelD dock I pet1bb, C cell. Trx-dock pet32, C cell. cohesin pet32, C therm CipA Coh I pet21b, C therm CohII pQE30 , CipA Dockerin C therm pQE30,pCip7 (Coh7) C therm pQE30 ,CelD Dock C therm pQE30, C therm dock II pQE30  
24/08/2012MiniprepsKevinsee above  
27/08/2012Heat shock transformationPhillipLinB pDE5TI7  
30/08/2012Liquid culturesKevinnFF GFP J04500 in DH5a, XL1 Blue, TOP10  
9/1/2012PCR amplification of D3 domainKevin   
01/09/2012PCRKevinRv2579, Coh2, Doc2, xDoc2  
01/09/2012GelKevinxylE XS  
01/09/2012GelKevinD3, GE LinB, GE xylE XS  
03/09/2012PCR OEKevinLinB, CohII, Rv2579, D3  
03/09/2012Digestion with DpnIPhillipLinB, CohII, Rv2579, D3  
03/09/2012MiniprepKevinlinB, CohII, Rv2579, D3, DH5a + J04500  
05/09/2012PCR of RFPDavid   
05/09/2012Digestion with XPPhillipMPP LinB, rv2579, CohII, D3 full constructs, DH5a + J04500  
05/09/2012PCRPhillipsee above  
05/09/2012PCRPhillipmgfp-5  
14/09/2012Digestion with EPKevinJ04500, RFP FC J04500, CohII FC, D3 FC, Rv2579 FC, LinB FC  
15/09/2012Liquid culture and miniprepKevinxylE FC  
15/09/2012GelKevinRFP, xylE FC, D GFP, D mgfp, D CohII, D xylE, D LinB, Rv2579  
15/09/2012LigationKevinD3 domain into psb1c3  
16/09/2012GelBeinimgfp, xdoc, doc, xylE FC EP GE, D RFP GE, D mgfp, D cohII, D linB, D Rv2579  
16/09/2012Digestion with SPBeiniMPP DH5a + J04500, RBS B0034 enzymatic purification using Biobasic kit
16/09/2012Liquid cultures and miniprepKevin, PhillipD3 FC Ligation trials  
18/09/2012LigationsKevinxylE FC GE and RFP FC GE into pSB1C3  
18/09/2012PCR overlap extensionPhillipD RFP, D GFP, D mgfp, D cohII, D linB, D Rv2579 (all GEs)  
18/09/2012Heat shock transformation of overlap extensions and ligationsKevinsee above  
19/09/2012Liquid culturesKevinlig RFP, D3 RFP OE, D3 GFP OE, D3 linB OE, D3 Rv2579, D3 mgfp OE, D3 cohII OE, D3 ctrl  
21/09/2012MiniprepsAaron, Beinisee above  
22/09/2012PCR OE of RFP with dockerin IVictorRFP, dockerin I7 trials with varying ratiosused sec tag left primer and dock right primer
22/09/2012Heat shock transformationBeinixylE FC AK into XL1 Blue  
23/09/2012GelFaisalxylE, cohII, linB, Rv2579, mgfp, D3 ctrl, sxdoc, sdoc  
23/09/2012PCRKevinsdoc and XS RFP doc with sec tag 2 primer  
23/09/2012DigestionsBeinisxdoc XP, xylE with SN, xylE with EP, CohII with SP, mgfp with EP  
23/09/2012GelPhillipsee above  
24/09/2012Heat shock transformationKevinJ04450, RFP D3 OE  
24/09/2012Glycerol stockKevinxylE full fliC deletion construct in pSB1C3 in XL1 Blue  
24/09/2012250mL cultureKevinxylE FC pSB1C3 XL1 Blue  
25/09/2012GelBeinigel extraction sxdoc RFP XP, sdoc RFP XP, xylE EP, MGFP D3 fliC EP  
25/09/2012Heat shock transformationBeiniMPP D3 FC lig into XL1 Blue, MPP D3 CohII OE into XL1 Blue  
25/09/2012LigationsKevinsdoc RFP J04500, sxdoc RFP J04500, mgfp pSB1C3  
26/09/2012Liquid culturesBeiniD3 FC, xylE FC AK, cohII FliC, mgfp5 FliC, sdoc RFP b0034, sxdoc RFP B0034  
26/09/2012MiniprepPhillipsee above  
29/09/2012Heat shock transformationsKevinD3 FliC, D3 LinB FliC, D3 Rv2579 FliC, D3 Coh FliC, D3 RFP FliC, RFP deletion FliC, J04450, mgfp5 FliC transformed into TOP10
30/09/2012Liquid culturesBeinisee above  
02/10/2012Protein isolationKevinmgfp5 FliC, J04450, D3 FliC, D3 LinB, RFP deletion FliC, D3 Rv2579 FliC, D3 RFP FliC, xylE deletion FliC, D3 CohII FliC5 mL cuturesheated 65C for 15 mins
02/10/2012xylE assayKevin   
03/10/2012Flagella isolation precedureKevin successful pellet in all samples

Given that the vast majority of our work now is in the lab, we will only be updating the "Labwork" section of the Notebook.

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