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| | <a href="https://2018.igem.org/Team:Bio_Without_Borders/LabBook">Lab Book</a> | | <a href="https://2018.igem.org/Team:Bio_Without_Borders/LabBook">Lab Book</a> |
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| − | <dt><b><font color="#009999">Week 1 (June 4-8)</font></b></dt>
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| − | <dd>•Made competent cells and aliquoted them into 25 1.5 mL tubes (50 microL each)</dd>
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| − | <dt><b><font color="#009999">Week 2 (June 11-15)</font></b></dt>
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| − | <dd>•Testing competent cells using iGem DNA transformation samples.</dd>
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| − | <dd>• Tested to see if they grew better on a freezer block versus an ice bath. They grow better in an ice bath.</dd>
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| − | <dd>• Ran colony PCR and plated bacteria on antimicrobial plate. </dd>
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| − | <dd>• cPCR, chloramphenicol, and culturing protocols</dd>
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| − | <dd>• Ran 1% agarose gel for cPCR</dd>
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| − | <dd>• Plasmid prep of pSB1C3</dd>
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| − | <dt><b><font color="#009999">Week 3 (June 18-22)</font></b></dt>
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| − | <dd>•Made chloramphenicol LB plates</dd>
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| − | <dd>•Restriction enzyme digest of iGem plasmid pSB1C3 using EcoR1 and Pst1</dd>
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| − | <dd>•Made digest master mix; ran an ezyme digest of iGem plasmids pSB1A3 and pSB1K3
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| − | Performed ligation of </dd>
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| − | <dd>pSB1A3 and pSB1K3; transformed pSB1A3 in competent cells on ampicillin plate</dd>
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| − | <dd>•Created a 50mg/mL kanamycin solution and then aliquoted it to about 80 1.5mL tubes. </dd>
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| − | <dd>•Inoculated LB with ampicillin colonies (with insert).</dd>
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| − | <dd>•Streaked ampicillin (1A3) colony onto new plates.</dd>
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| − | <dd>•Conducted cPCR for 1A3 colonies with insert.</dd>
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| − | <dd>•Filled out first draft of safety form</dd>
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| − | <dd>•Went to Brighton Beach to examine horseshoe crabs' habitat and found a part of the carcass</dd>
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| − | <dt><b><font color="#009999">Week 4 (June 25-29)</font></b></dt>
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| − | <dd>•Conducted cPCR for pSB1K3.</dd>
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| − | <dd>•Ran 1% agarose gel of pSB1A3 with the insert (colonies that were re-streaked).</dd>
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| − | <dd>•Ran 1% agarose gel of pSB1K3 with insert.</dd>
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| − | <dd>•Cleaned laboratory material.</dd>
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| − | <dd>•Set up pSB1C3 backbone amplification PCR using Q5.</dd>
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| − | <dd>•Ran gel of pSB1C3 j04450 insert with Q5 PCR to confirm size.</dd>
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| − | <dd>•Amplification of pSB1C3 j04450 insert with Phusion PCR (because Q5 did not work). Confirmed size with gel electrophoresis.</dd>
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| − | <dd>•Plasmid prep of pSB1A3</dd>
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| − | <dd>•Digest of amplified pSB1C3 j04450 insert (Phusion PCR).</dd>
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| − | <dd>•Amplified linearized backbone from pSB1C3 and pSB1A3 with Phusion PCR</dd>
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| − | <dd>•Ran a gel for pSB1A3 and pSB1C3 plasmid prep from day before to check band size</dd>
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| − | <dd>•Purified (accidentally-amplified with Phusion) PSB1C3 j04450 insert, as well as the circular (made into linear) pSB1A3 and pSB1C3 backbones with PCR clean-up</dd>
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| − | <dt><b><font color="#009999">Week 5 (July 2-9)</font></b></dt>
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| − | <dd>•Went to Jones Beach to examine horseshoe crabs' habitat and found a carcass</dd>
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| − | <dd>•Worked on GoFundMe campaign page</dd>
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| − | <dd>•Digested pSB1K3 backbone, ligated it with the J04450 insert (from purified J04450 insert from 6/27), and transformed cells for Kanamycin resistance & plated them</dd>
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| − | <dd>•Ran cPCR for transformed kanamycin resistant colonies</dd>
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| − | <dd>•Coded format for iGEM website</dd>
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| − | <dd>•Inoculated LB broth with transformed kanamycin resistant colonies</dd>
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| − | <dd>•Made iGEM Facebook page</dd>
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| − | <dt><b><font color="#009999">Week 6 (July 7-13)</font></b></dt>
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| − | <dd>•Human practices: PCR and Pizza.</dd>
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| − | <dd>•Ran cPCR of pSB1K3 ith j04450 insert.</dd>
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| − | <dd>•Re-did cPCR of pSB1K3 and ran gel (failed again).</dd>
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| − | <dd>•Made chloramphenicol solution, as well as chloramphenicol plates.</dd>
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| − | <dd>•Transformed the cellulose binding domain (a.k.a. BBa_K1478001) from distribution plate into competent cells and plated them on chloramphenicol.</dd>
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| − | <dd> (We need to figure out which primers to so that we can observe a band from cPCR of pSB1K3).</dd>
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| − | <dd>• Inoculated LB broth with transformed cellulose binding domain containing colonies</dd>
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| − | <dd>• Made new VF2 and VR primers and performed cPCR with them for BBa_K1478001 and pSB1K3</dd>
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| − | <dd>• Ran gel for both pSB1K3 and BBa_K1478001 (failed again). Maybe we order new primers and change PCR master mix.</dd>
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| − | <dd>• Resuspended from distribution and transformed BBa_E1010(RFP,chloramphenicol backbone), BBa_K648013(GFP, chloramphenicol backbone) and BBa_E0040(GFP, ampicillin backbone) in competent cells.</dd>
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| − | <dd>• Plasmid prep of two pSB1K3 samples and two of cellulose binding domain BBa_K1478001 samples.</dd>
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| − | <dd>• Plated all transformation cells</dd>
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| − | <dt><b><font color="#009999">Week 7 (July 16-20)</font></b></dt>
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| − | <dd>• BBa_E1010(RFP,chloramphenicol backbone) and BBa_K648013(GFP, chloramphenicol backbone) transformed plates did not work. BBa_E0040(GFP, ampicillin backbone) transformed plate had two colonies present.</dd>
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| − | <dd>• cPCR was conducted for two BBa_E0040 (GFP) transformed colonies.</dd>
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| − | <dd>• Ran gel of digested backbone of BBa_E0040. (7/17)</dd>
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| − | <dd>• Made plates of LB with chloramphenicol.</dd>
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| − | <dd>• Performed transformation with 1uL of E1010(RFP) and K648013(GFP), on chloramphenicol. Result: There were no colonies on plates with E1010(RFP),and approximately 3 or 4 colonies grew on plate with K648013(GFP).</dd>
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