View source for Team:ETH Zurich/Notebook/Text

22/06/2015
<p>
	<ul>
	<li>Interlab study:ligation and transformation</li>
	<li>Redo transformation that failed</li>
	<li>Order primers, Antibodies and lactate kits</li>
	</ul>
</p>

23/06/2015
<p>
	<ul>
	<li>Send trafos (bricks and Interlab parts) for sequencing</li>
	<li>FACS training</li>
	<li>Colony PRC for trafos</li>
	<li>Make cryostocks of interlab positive and negative control</li>
	<li>Design and order lldRO versions (incl lacO)
</li>
	<li>transform again InterLab ligated constructs (twice each)
</li>
	</ul>
</p>

24/06/2015
<p>
	<ul>
	<li>Colony PCR for InterLab constructs</li>
	<li>Send InterLab constructs for sequencing</li>
	<li>Annexin V part : put parts together with backbone
</li>
	<li>Amplify lldR from the genome
</li>
	<li>Make overnight culture for GFP cells
</li>
	<li>Make cryostocks of the 3 constructs (3 replicates)
</li>
	<li>Retransform InterLab control I20270?</li>
<li>Design and order all remaining constructs/primers</li>
	</ul>
</p>

25/06/2015
<p>
	<ul>
	<li>Got labelled annexin</li>
	<li>ON culture for I35104 and the three InterLab promoters (15mL in total) from picked colony
</li>
	<li>Run lldR PCR on gel and purify the construct!
</li>
	<li> Look into sTRAIL handling (prepare for use)
</li>
	<li>GFP competent cells</li>
	</ul>
</p>

26/06/2015
<p>
	<ul>
	<li>Miniprep Of InterLab study</li>
	<li>Digestion of InterLab-purification</li>
	<li>lldP amplification from the genome--> didn't work!</li>
	<li>Amplify INP from the biobrick and purify from gel</li>
	<li>Alliquoting of TRAIL</li>
	</ul>
</p>

29/06/2015
<p>
	<ul>
	<li>Throw away the incorrect parts</li>
	<li>Transform TOP10 and gfp strains for efficiency study</li>
	<li>Measure concentration of fG0008-fG0012</li>
	<li>Ligate InterLab fragments</li>
	<li>InterLab transformation</li>
	<li>Get Omp-8 cells</li>
	<li>TOP10 ON culture</li>
	</ul>
</p>

30/06/2015
<p>
	<ul>
	<li> Make TOP10 cryostock</li>
	<li>Transformation efficiency study</li>
	<li>Colony PCR Interlab</li>
	<li>Interlab sent to sequencing</li>
	<li>Measure conc of lldP</li>
	<li>Transform pSEVA</li>
	<li>Make new LB+CM plates</li>
	<li>Make an ON culture from c13</li>
	</ul>
</p>


01/07/2015
<p>
	<ul>
	<li>Passage cells </li>
	<li>Pick up Omp-8</li>
	<li>Use B0032 (sequenced) for negative control of InterLab</li>
	<li>Make new LB</li>
	<li>Colony PCR</li>
	<li>Send device to sequencing</li>
	<li>Omp-8 ON culture at 25ºC</li>
	<li>pSEVA ON culture</li>
	<li>Competent cells</li>
	</ul>
</p>

02/07/2015
<p>
	<ul>
	<li>Miniprep pSEVA</li>
	<li>Send one sample of pSEVA for sequencing</li>
	<li>Repeat interlab 2 colony PCR (all negative..)</li>
	<li>Depending on sequencing results: throw out or correctly label the cryo tube which is now called p32-1</li>
	<li>Sample resubmitted for sequencing with reverse primer oiG002</li>
	<li>Remake Omp-8 preculture</li>
	<li>Make TBF buffer</li>
	</ul>
</p>

03/07/2015
<p>
	<ul>
	<li>Mammalian cell passaging</li>
	<li>Order oligos</li>
	<li>Amplification of annexinV</li>
	<li>Digestion of pSEVA low copy</li>
	<li>TPR digestion: failed partly</li>
	<li>Digest of InterLab 2</li>
	</ul>
</p>

06/07/2015
<p>
	<ul>
	<li>Digest pSEVA371</li>
	<li>SOC medium</li>
	<li>Gibson assembly annexin</li>
	<li>Plan mammalian experiments with TRAIL</li>
	<li>Transform interlab, annexinV-pSEVA, pSEVA371, pSEVA271</li>
	<li>Overnight culture of Omp-8 at 16°, 25°, 37°</li>
	<li>Digest pSEVA371</li>
	</ul>
</p>

07/07/2015
<p>
	<ul>
	<li>Passage cells</li>
	<li>Asked Tania</li>
	<li>Comment with Margaux the mammalian experiments</li>
	<li>Cryostocks</li>
	<li>Primer design</li>
	<li>Colombia -> protocol sent</li>
	<li>PCR for lldP-lldR</li>
	</ul>
</p>

08/07/2015
<p>
	<ul>
	<li>Make LB-agar and LB-medium</li>
	<li>pSEVA miniprep</li>
	<li>pSEVA digest and purification with miniprep</li>
	<li>Purify lldR-lldP fragments (fridge) (digest the template first)</li>
	<li>PCR for remaining INP-Annexin fragments</li>
	<li>2% agarose gel for remaining INP-Annexin fragments</li>
	<li>Make ON cultures of all useful BioBrick transformants for cryo stocks</li>
	</ul>
</p>

09/07/2015
<p>
	<ul>
	<li>Start mammalian experiments+ TRAIL
</li>
	<li>Shopping list </li>
	<li>Colony PCR annexinV colony
</li>
	<li>Competent Omp-8
</li>
	<li>Mammalian cell FACS</li>
	<li>Gibson assembly</li>
	<li>Transformation INP-An, lldP-lldR, AnV, p27 and p30</li>
	<li>Make LB</li>
	<li>Preparation of interlab study</li>
	</ul>
</p>

10/07/2015
<p>
	<ul>
	<li>Colony PCR</li>
	<li>Cryostocks (still missing 27 and 30)</li>
	<li>InterLab FACS done!</li>
	<li>Sequencing</li>
	<li>Make ITA buffer</li>
	</ul>
</p>

13/07/2015
<p>
	<ul>
	<li>Compare results: no results</li>
	<li>Colony PCR of lldR-lldP</li>
	<li>GA for lldR-lldP diluted</li>
	<li>pSEVA271 digestion and miniprep</li>
	<li>Preparation of lldR-lldP plates</li>
	<li>After meeting planning of experiments</li>
	<li>ON culture of AnV and INP-AnV</li>
	</ul>
</p>

14/07/2015
<p>
	<ul>
	<li>Cryostock of interlab device 2</li>
	<li> lldR-lldP colony PCR</li>
	<li>Miniprep of ON of AnV and INP-AnV, send to sequence</li>
	<li>TPR re-do</li>
	<li>BioBrick assembly of luxI-term</li>
	<li>Gel-purify all PCR products from monday</li>
	<li>Evening: set up mammalian cells with christian</li>
	<li>pSEVA ON for cryostock</li>
	<li>BB assembly ligations overnight</li>
	</ul>
</p>

15/07/2015
<p>
	<ul>
	<li>Start TRAIL and E Coli mammalian experiments, monitor all day</li>
	<li>Colony PCR for new colonies of AnV</li>
	<li>ON cultures of AnV</li>
	<li>lldR-lldP miniprep</li>
	<li>Redo digest for luxI-term assembly (C0161)</li>
	<li>Transformation of bb-assembled stuff</li>
	<li>Send for sequencing: the "lldR-lldP" plasmid (once with oiG0003 and once with oiG0004)</li>
	<li>Make overnight culture from cryostock of INP-Annexin_2 </li>
	<li>ON of GFP cells for co-culture</li>
	<li>Gibson Assemble Plasmids p62-81</li>
	<li>Transformation of gibson assembled plasmids</li>
	</ul>
</p>

16/07/2015
<p>
	<ul>
	<li>Make cultures of BB assembled, colony PCR</li>
	<li>Make cultures of p62-81, colony PCR, make overnight cultures</li>
	<li>Pick up sequencing labes from shop- afternoon (let grow for a long time): miniprep a lot of INP-Annexin, send for sequencing with oiG0004</li>
	<li>Omp-8 transformation efficiency</li>
	<li>Set up mammalian cells and bacteria for lactate experiment</li>
	<li>Mini-prep C0161 and B0012</li>
	<li>Digest C0161 and B0012</li>
<li>Overnight culture for interlab</li>
	</ul>
</p>

17/07/2015
<p>
	<ul>
	<li>Set up reader plate</li>
	<li>Gibson assembly for lldR w/o lldP</li>
	<li>Transform and plate lldR constructs</li>
	<li>Lactate experiment: change medium at 12:30</li>
	<li>Make cryostocks of all things to sequence (annexin and all assemblies)</li>
	<li>Send all things for sequencing </li>
	<li>Use TOP10 culture for lactate experiment</li>
	<li>Repeat co-culture 14:00-16:00</li>
	<li>Redo digest of C0161 and B0012 (includde test of enzymes)</li>
	</ul>
</p>

19/07/2015
<p>
	<ul>
	<li>Set up overnight cultures of cryos made on friday plus maybe more of each assembly from 0715 plus from assembly on 0717</li>
	</ul>
</p>
 

20/07/2015
<p>
	<ul>
	<li>Check sequencing results</li>
	<li>Data process: lactate</li>
	<li>Redo digest to test enzymes</li>
	<li>Send stuff to sequence</li>
	<li>New fragments with new fragments</li>
	<li>Make colony PCR</li>
	<li>Throw away stuff from cryostocks</li>
	<li>Make overnight cultures of what is missing: (p72 and p73)</li>
	<li>Plan mammalian experiments to know how many cells we need each day
</li>
	<li>Process data (FACS, InterLab and lactate)
</li>
	<li>Make o/n cultures for interlab in triplicates
</li>
	</ul>
</p>

21/07/2015
<p>
	<ul>
	<li>Early do interlab and InterLab sumitted</li>
	<li>Keep making new fragments with new primers</li>
	<li>If sequencing is correct, make fragments for PL(5)</li>
	<li>Send p72 to sequence</li>
	<li>New fragments: DPN1 digest, gel, purify from gel</li>
	<li>Miniprep stuff</li>
	<li>Continue TPR, ligate and transform</li>
	</ul>
</p>

22/07/2015
<p>
	<ul>
	<li>TRAIL experiment start at 9:00 fro 3T3</li>
	<li>Repeat transformation of lldR and TPR</li>
	<li>Miniprep c44 and c45 and INP-Ann and p72 to have more plasmid
</li>
	<li>Sample organisation study
</li>
	<li>Colony PCR and make ON culture of lldR andTRP(12_114_34)</li>
	<li>Send 50 for sequencing with different primer</li>
	<li>Miniprep p73 variants for sequencing</li>
	<li>Colony PCR</li>
	<li>TRAIL experiment 15:30-17:00 FACS machine</li>
	<li>Continue TPR assembly
</li>
	<li>Digest BioBrick backbone
</li>
	<li>Annexin buffer binding made new</li>
	<li>TOP10 ON</li>
	</ul>
</p>

23/07/2015
<p>
	<ul>
	<li>Colony PCR of GFP-INP-Annexin</li>
	<li>Colony PCR of lldR, 12_117, TPR</li>
	<li>Make more LB</li>
	<li>Digestion Annexin- bb--> no band on annexin</li>
	<li>Send TPR for sequencing</li>
	<li>ON culture of AnnexinV and B0012</li>
	<li>Making Competent Top10 cells</li>
	</ul>
</p>

24/07/2015
<p>
	<ul>
	<li>Miniprep annexin</li>
	<li>Digest INP-An, B0012</li>
	<li>Retransform TPR minipreped yesterday</li>
	<li>Amplify TRP to use in assemblies if sequencing is correct</li>
	<li>Send a lot of things for sequencing (lldR, 12_117)
</li>
	<li>Do gibson assembly for pl5, transform</li>
	</ul>
</p>
26/07/2015
<p>
	<ul>
	<li>Precultures</li>
	</ul>
</p>
27/07/2015
<p>
	<ul>
	<li>Experiment with Jurkat cells started (lactate)</li>
	<li>Ligation AnV-BB</li>
	<li>Ordered new lldR-lldP plasmid gblocks</li>
	<li>Miniprep ON cultures (TPR and PL(5))</li>
	<li>Send ON cultures (TPR and PL(5))for sequencing</li>
	</ul>
</p>

28/07/2015
<p>
    <ul>
    <li>Make more TAE</li>
    <li>Miniprep lldR ON cultures</li>
    <li>Send lldR for sequencing</li>
    <li>lldR gibson assembly</li>
    <li>lldR transformation</li>
    <li>ON cultures of AnV(c73), INP-AnV(c55), PL(5)(cryos 2b, 1, 2a),
PL(7), TOP10 cells with no plasmid (c13)</li>
    </ul>
</p>

29/07/2015
<p>
    <ul>
    <li>Book plate reader</li>
    <li>Think about PS-coated</li>
    <li>ON Cultures annexin in BBBB</li>
    <li>Experiment Annexin-INP in the membrane</li>
    <li>Order one TPR in IDT</li>
    </ul>
</p>

30/07/2015
<p>
    <ul>
    <li>Digest of INP-Annexin (failed)</li>
    <li>Digest of luxI and B0012 (failed)</li>
    <li>ON cultures pSEVA 271 and 371</li>
    <li>ON cultures of B0012 (c27), INP-AnV, luxI</li>
    <li>AnV in BioBrick BackBone sent to sequencing</li>
    <li>Western blot for annexin-inp</li>
    <li>Make ON cultures from p101 (lldR-lldP) plates</li>
    <li>Make new LB and LB-agar</li>
    <li>ON cultures of PL(6)</li>
    <li>Mammalian set up</li>
    </ul>
</p>

31/07/2015
<p>
    <ul>
    <li>Miniprep pSEVA</li>
    <li>Miniprep PL6</li>
    <li>Miniprep and digest of B0012 (c27), INP-AnV and luxI and ligate</li>
    <li>TRAIL jurkat and 3t3</li>
    <li>Lactate all lines + calibration curve</li>
    <li>PCR for fragments 74-80, 97, 98</li>
    <li>Digest test</li>
    </ul>
</p>

01/08/2015
<p>
    <ul>
    <li>Gibson assembly lldP-lldR</li>
    <li>Transformation</li>
    </ul>
</p>

02/08/2015
<p>
    <ul>
    <li>Colony PCR of up to 20 colonies with primers oi3 and oi19</li>
    <li>Make ON cultures, two of every positive colony from the PCR</li>
    </ul>
</p>

03/08/2015
<p>
    <ul>
    <li>Order gblock</li>
    <li>Track selection</li>
    <li>Cryostocks and miniprep lldR-lldP</li>
    <li>Sent stuff for sequencing</li>
    <li>Lactate evaluation (FAIL)</li>
    <li>Try PCR (p24 + o61/o62 = f64) with Q5 2x Master mix (783bp fragment) at 62, 64 and 66°C (FAIL)</li>
    <li>CRYO STOCKS!!!! miniprep and send for sequencing: lldR-lldP</li>
    <li>4pm meeting doctor</li>
<li>Order primers</li>
    </ul>
</p>

04/08/2015
<p>
    <ul>
    <li>Start cultures of lldR and lldR-lldP for electrocompetent cells (let grow to OD 0.6)</li>
    <li>Prepare everything for lactate based experiments</li>
    <li>cultures of backbone 371</li>
    <li>order cryotubes</li>
    <li>transform pl3 into intermediate plasmid strain</li>
    <li>make gibson assembly again and transform</li>
    </ul>
</p>

05/08/2015
<p>
    <ul>
    <li>Crystock of pSEVA</li>
    <li>Miniprep pSEVA+ failed digest</li>
    <li>Colony PCR of PL3</li>
    <li>GA for lldP-lldR (old-old one-with low concentration-): one 1h 45ºC, the other 2h 50ºC</li>
    <li>Seeding new mammalian cell line</li>
    <li>ON culture of pSEVA</li>
    <li>Redo Gibson Assembly of p64 and p73, transform and incubate o/n</li>
    </ul>
</p>

07/08/2015
<p>
    <ul>
    <li>Miniprep p50</li>
    <li>Assemble lldPR, PL3, PL4, PL5, INP-an and transformation</li>
    <li>Change concentrations and repeat the experiment of the transformants with PL3 and lldR</li>
    </ul>
</p>

08/08/2015
<p>
    <ul>
    <li>Colony PCR of all Trafos from Friday</li>
    <li>Redo assemblies of INP-AnV and PL(5)</li>
    <li>ON of INP-AnV and PL(5) from assemblies from friday</li>
    <li>Transformation of assemblies</li>
    </ul>
</p>

09/08/2015
<p>
    <ul>
    <li>Redo assemblies of PL(3) and PL(4)</li>
    <li>Trafo of assemblies</li>
    <li>Miniprep ON from Saturday</li>
    <li>Make ON of c89-c98</li>
    </ul>
</p>

10/08/2015
<p>
    <ul>
    <li>Colony PCR of PL(3) and PL(4)</li>
    <li>Make ON cultures of PL(3) and PL(4)</li>
    <li>Send things for sequencing (INP-AnV and PL(5))</li>
    <li>Assemble PL(9)</li>
    <li>Transform assembly of PL(9)</li>
    <li>Lactate calibration curve</li>
    <li>Lactate experiment mammalian cells</li>
    <li>Data evaluation</li>
    <li>Overnight cultures of INP-annexins and c89-c98</li>
    <li>Lactate experiment e coli</li>
    </ul>
</p>

11/08/2015
<p>
    <ul>
    <li>Cryostocks PL3, Pl4, lldr, miniprep, sent for sequencing</li>
    <li>Colony PCR PL9</li>
    <li>Lactate experiment with bacteria twice</li>
    <li>ON of pseva271</li>
    <li>Colony PCR of PL(9), make ON if it worked (didn't work)</li>
    <li>Throw away not confirmed cryos</li>
    <li>Redo PL9 assembly and trafo</li>
    <li>Do new assemblies with just arrived primers</li>
    </ul>
</p>

12/08/2015
<p>
    <ul>
    <li>Miniprep and digest bb</li>
    <li>PL4 and PL3-3 sent for sequencing</li>
    <li>Colony PCR of PL(9), didn't work</li>
    <li>Redo assembly for PL(9) with different backbones and transform</li>
    <li>Assemble and transform lldR-lldP, annexinseva, annexinbb, inpyfp</li>
    <li>TRAIL experiment</li>
    <li>Lactate addition experiment with bacteria twice</li>
    </ul>
</p>


13/08/2015
<p>
    <ul>
    <li>Colony PCR</li>
    <li>Replan mammalian cell experiment: why TRAIL is not working?</li>
    </ul>
</p>

14/08/2015
<p>
    <ul>
    <li>Cryostocks of new cultures</li>
    <li>Colony PCR of PL9</li>
    <li>Miniprep of lldrp, pl9, pl3, pl4 and AnnBBBB and sent some for sequencing</li>
    <li>Miniprep of lldrp, pl9, pl3, pl4 and AnnBBBB and sent some for sequencing</li>
    <li>Preparation of apoptotic cells (TRAIL)</li>
    <li>Co-culture HL-60 and bacteria</li>
    <li>Split cells</li>
    </ul>
</p>

15/08/2015
<p>
    <ul>
    <li>Lactate dose-response experiment with bacteria</li>
    <li>Darmstadt visit</li>
    </ul>
</p>

16/08/2015
<p>
<ul>
    <li>Lactate dose-response experiment with bacteria</li>
    <li>Darmstadt visit</li>
    </ul>
</p>

17/08/2015
<p>
    <ul>
    <li>Send for sequencing lldrp, pl9 etc</li>
    <li>Good ones PL3 measured at high conc</li>
    <li>Bad ones (65-68) PL(3) measured at same conc</li>
    <li>Microscopy TRAIL</li>
    <li>Miniprep</li>
    <li>Repeat lactate measurement for cells at 6h</li>
    <li>Make overnight cultures of PL(3) versions, top10, lldr, lldplldr</li>
    </ul>
</p>

18/08/2015
<p>
    <ul>
    <li>Bad ones of PL(3) in plate reader at 25mM, 6.3, 3.1, 390uM, 100uM, 10uM</li>
    <li>Make fragments from PL(9)</li>
    <li>Measure lactate from mammalian cells from yesterday (morning before other exp.)</li>
    <li>Make competent cells</li>
    <li>Gp62, p69, p70 and p71 again with high concentration of lactate at gain 70</li>
    <li>More minipreps</li>
    </ul>
</p>

19/08/2015
<p>
    <ul>
    <li>p68, p66, p67 and p63 for all concentrations of lactate </li>
    <li>Send things for sequencing</li>
    <li>Remake fragment f48 failed</li>
    </ul>
</p>

20/08/2015
<p>
    <ul>
    <li>Make SOC medium</li>
    <li>ON cultures pSEVA and annexinV, PL5, INP AnV and AnnexinV in BBBB</li>
    <li>Transform PL4 in lacI</li>
    <li>Remake fragment f48 and reassemble/retransform PL(4)a</li>
    <li>Maybe redo good plasmids two per plate whole range (all)</li>
    <li>Double transform PL(4) into lacI strain (c29)</li>
    <li>Double trafo if sequencing is good (PL(4) and PL(3) versions into lldPlldR)</li>
    </ul>
</p>

21/08/2015
<p>
    <ul>
    <li>Colony PCR of double transformants and PL4</li>
    <li>Miniprep pSEVA271 and digest (133ng/uL)</li>
    <li>Western blot to check annexin presence</li>
    <li>Buy enzymes for GA</li>
    </ul>
</p>
22/08/2015
<p>
    <ul>
    <li>GA buffer</li>
    <li>Gels for PCR</li>
    <li>Cryostocks of double transformants</li>
    <li>Split mammalian cells </li>
    <li>Picture of western blot</li>
    <li>Fragments lldR-lldP--> didn't work. Plan again. </li>
    <li>Miniprep PL4--> not enough concentration for sequencing. Wait until new kit arrives</li>
    </ul>
</p>
23/08/2015
<p>
    <ul>
    <li>Western blot for AnnexinV in BBBB and pSEVA</li>
    <li>Lactate experiment with p75 and p77</li>
    </ul>
</p>


24/08/2015
<p>
    <ul>
    <li>Take lldpr operon from genome with oiG12 and oiG19. use phusion PCR with 5 min heating in the beginning from a resuspended colony or from cryotube, best is empty TOP10.</li>
    <li>Western blot with higher concentration of protein.</li>
    <li>Split cells</li>
    </ul>
</p>

25/08/2015
<p>
    <ul>
    <li>Get beads with streptavidin</li>
    <li>Colony PCR lldP-lldR (oiG76 and oiG75) and make cultures</li>
    <li>Check 3T3 cells and split </li>
    <li>Digest B0012 for obtaining BBBB</li>
    <li>INP-AnV, pl5 fragment</li>
    <li>Experiment with beads (microscopy and FACS): we have red fluorescence in the beads!</li>
    <li>Make LB</li>
    </ul>
</p>

26/08/2015
<p>
    <ul>
    <li>p69 with and without lactate experiment</li>
    <li>miniprep INP-Annexin, PL5, lldRP, PL4a colonies</li>
    <li>Send lldRP to sequence and test digest</li>
    <li>Digest, ligate and transform INP-Annexin and PL5</li>
    <li>Primer design</li>
    <li>Make agar</li>
    </ul>
</p>

27/08/2015
<p>
    <ul>
    <li>p62 and p70 whole range with and without lldR</li>
    <li>Split mammalian cells</li>
    <li>Miniprep more lldPR</li>
    <li>Colony PCR INP-Annexin and PL5 in BBBB</li>
    <li>Discuss presentation</li>
    <li>Make fragments for restriction plan for lldPR</li>
    <li>Purify from gel: amplifiaction of gblocks. fail.</li>
    <li>Overnight cultures</li>
    <li>Found HER2 cells</li>
    <li>Got superkillerTRAIL</li>
    <li>Double transformation of LP3 and PL4 versions into lldPR if sequencing is correct</li>
    </ul>
</p>

28/08/2015
<p>
    <ul>
    <li>Beads and antibody experiment for everything related to annexin (incl from BBBB)</li>
    <li>Miniprep p68</li>
    <li>Colony PCRs for double trafos</li>
    <li>TRAIL 2h and 4h with Jurkat and HL60. use bacteria to detect</li>
    <li>Add PL(5) cells to mammalian cells after TRAIL</li>
    <li>Purify fragemtn for restriction plan</li>
    <li>Digest and ligate lldPR restriction plan</li>
    <li>Transform lldPR restriction plan</li>
    <li>Test PL9 response to AHL</li>
    <li>Test p71+-lldR for whole conc range</li>
    <li>Mini prep and send for sequencing: PL5 and INP-Annexin in BBBB</li>
    </ul>
</p>

29/08/2015
<p>
    <ul>
    <li>Make fragments for GA</li>
    <li>Colony PCR reeated for p71+lldPR</li>
    <li>Colony PCR for lldPR restriction plan</li>
    <li>Make cryos of all double trafos with lldPR</li>
    <li>Make overnight cultures of all double trafos with lldPR and do GA with them</li>
    <li>Set up first presentation draft</li>
    <li>Set up PCR for PL12 fragment</li>
    <li>Digest more backbone</li>
    </ul>
</p>


30/08/2015
<p>
    <ul>
    <li>Got the cryos for Stockholm</li>
    <li>Digest more backbone</li>
    <li>Colony PCR lldRPres, PL1, PL10</li>
    </ul>
</p>

31/08/2015
<p>
    <ul>
    <li>Pick up stuff from shop</li>
    <li>lldP-lldR with restriction plan, PL1 and PL10 sent to sequence</li>
    <li>Digestion of backbone</li>
    <li>Colony PCR --> nothing is working again </li>
    <li>GA of PL10 and PL1</li>
    <li>Lactate experiment </li>
    <li>Pick up and thaw SKBR-3 cells </li>
    </ul>
</p>

01/09/2015
<p>
    <ul>
    <li>Colony PCR of p71</li>
    <li>Fragment purification for GA</li>
    <li>Mammalian lactate experiment</li>
    <li>Beads experiment: failed</li>
    <li>Take a look at lldP-lldR under the microscope (different shape)</li>
    <li>GA of PL10</li>
    </ul>
</p>

02/09/2015
<p>
    <ul>
    <li>Colony PCR of PL10--> nothing works (desperation)</li>
    <li>Lactate experiment with bacteria</li>
    <li>Measure lactate in mammalian cells: collect up to 24h</li>
    <li>Make overnight cultures</li>
    </ul>
</p>

03/09/2015
<p>
    <ul>
    <li>Colony PCR of PL12, and all versions of lldRPres and PL3 and PL4.</li>
    <li>Do the presentation for EPFL</li>
    <li>Human practices collaboration with EPFL</li>
    <li>Prepare fragments for PL2</li>
    <li>Analyze data for mammalian lactate experiment</li>
    <li>Prepare more samples for characterization of our promoters.</li>
    </ul>
</p>

04/09/2015
<p>
    <ul>
    <li>Purify fragments</li>
    <li>Split cells</li>
    <li>Set up new characterization experiment for lldRPres</li>
    <li>ON cultures for lldRP experiment</li>
    </ul>
</p>

05/09/2015
<p>
    <ul>
    <li>Chip experiment</li>
    <li>Experiment to see green fluorescence in minimal medium</li>
    <li>Amplification and purification of fragments for BioBrick</li>
    <li>Lactate experiment</li>
    <li>ON cultures of bacteria sent by Stockhoml, lacI, p76, p74, p73 and TOP10.</li>
    </ul>
</p>

06/09/2015
<p>
    <ul>
    <li>Assembly and transformation of promoters in biobrick</li>
    <li>Lactate experiment</li>
    <li>Transform GFP into Stockholm cells and fragments into BBBB</li>
    <li>Split cells</li>
    <li>ON cultures of PL5 in BBBB, PL12, TOP10</li>
    <li>Colony PCR</li>
    </ul>
</p>

07/09/2015
<p>
    <ul>
    <li>Miniprep and send for sequencing: PL2, PL12, PL10</li>
    <li>ON cultures of PL12, PL5, RFP, PL10, TOP10</li>
    <li>Repeat fragments</li>
    </ul>
</p>

08/09/2015
<p>
    <ul>
    <li>Beads experiment with PS and Ab</li>
    <li>Competent cells</li>
    <li>Lactate experiment for Jurkat and SK-BR-3</li>
    <li>Traansform Stockholm cells with GFP again</li>
    </ul>
</p>

09/09/2015
<p>
    <ul>
    <li>Colony PCR of things in BioBrick BackBone</li>
    <li>Do new assemblies in BioBrick, previous didn't work</li>
    <li>ON cultures</li>
    </ul>
</p>

10/09/2015
<p>
    <ul>
    <li>Preparation of fragments, GA of PL2 and transformation</li>
    <li>Colony PCR</li>
    <li>Digest of BioBrick BackBone. </li>
    <li>Ligation and transformation of parts for Registry (again)</li>
    <li>ON cultures</li>
    <li>Preparation of mammalian cells</li>
    </ul>
</p>

11/09/2015
<p>
    <ul>
    <li>Mammalian cells experiment</li>
    <li>Colony PCR of BBBB. Didn't seem to work...</li>
    <li>TRAIL experiment for concentration range + FACS</li>
    <li>Incorporate bacteria in apoptotic mammalian cells + FACS + microscopy</li>
    <li>Backbone cultures</li>
    <li>Introduce lldR and lldRP into bacteria</li>
    <li>Do Stockholm experiment</li>
    <li>ON cultures</li>
    </ul>
</p>

12/09/2015
<p>
    <ul>
    <li>Miniprep BB BB</li>
    <li>ON cultures</li>
    <li>Digest BB, digest material for biobrick</li>
    <li>Transform bacteria</li>
    </ul>
</p>

13/09/2015
<p>
    <ul>
    <li>Colony PCR of BBBB: some positives!</li>
    <li>Co-culture Jurkat and bacteria</li>
    <li>Induction of apoptosis and look in the microscope the result, with three different concentrations of Annexin Binding Buffer and in three temperatures.</li>
    <li>Prepare Stockholm cells</li>
    <li>Lactate measurements</li>
    </ul>
</p>

14/09/2015
<p>
    <ul>
    <li>Make fragments with newly arrived primers</li>
    <li>Colony PCR of biobricks: we have all the promoters now</li>
    <li>Stockholm experiment with mammalian cells</li>
    <li>Miniprep BBBB</li>
    </ul>
</p>

15/09/2015
<p>
    <ul>
    <li>TRAIL experiment: measure lactate concentration in apoptotic cells and test again ANnexin binding. --> no success</li>
    <li>Colony PCR of the backbone.</li>
    <li>ON cultures of lldP, lldPR in BB</li>
    <li>Co-culture HL-60 and bacteria</li>
    </ul>
</p>

16/09/2015
<p>
    <ul>
    <li>Colony PCR</li>
    <li>ON culture of pSEVA371, for 100mL</li>
    <li>Lactate measurement</li>
    <li>Last try for assembly of PL14 and PL1</li>
    </ul>
</p>

17/09/2015
<p>
    <ul>
    <li>Be sleep deprived</li>
    <li>Cry and panick</li>
    <li></li>
    <li></li>
    <li></li>
    <li></li>
    <li></li>
    <li></li>
    <li></li>
    </ul>
</p>