Map+of+Experiments+for+Phage+Library


 * gel extraction 6-2-09
 * PCANTAB5E 6-5-09
 * Transformation with X2 cut PCANTAB5E 6-8-09
 * RNA Isolation 6-9-09
 * Competent Cells 6-15-09
 * Transformation to check competent cells 6-17-09
 * phage display preparation 6-19-09
 * phage display 6-19-09
 * amplified eluate 6-23-09
 * rna extraction 6-30-09
 * transformation with scFv plasmid (primer calculations) 7-6-09
 * Spleen Extraction 7-8-09
 * RNA extraction 7-8-09
 * RNA gel 7-8-09
 * cDNA synthesis 7-9-09
 * cDNA gel 7-10-09
 * RNA extraction 7-11-09
 * synthetic scFv Library 7-12-09
 * synthetic library rescue 7-15-09 to 7-16-09
 * separating phage from cells 7-18-09
 * phage titer 7-18-09
 * phage titer results 7-19-09
 * phage titer calculations 7-20-09
 * helper plasmid (M13cp) Transformation 7-23-09
 * preparing electrically competent cells 7-24-09
 * m13 plasmid transformation 7-25-09
 * competent cells 7-26-09 to 7-27-09
 * m13 transformation with chloramphenicol 7-28-09
 * ER2738 Transformation 7-29-09
 * transformation of dh10b with pLACI 7-29-09
 * Lower concentration chloramphenicol 7-30-09
 * analyzing competent cell results 7-31-09
 * scFv plasmid transformation 7-31-09
 * transformation of helper plasmids 8-1-09
 * electrical competent cell preparation 8-2-09
 * Midi of M13cp 8-3-09
 * Electroporation of several plasmids 8/4/09
 * biotinylate ab 8/5/09
 * phage ELISA 8-5-09 to 8-6-09
 * Phage ELISA Results 8-7-09
 * Dot Blot Analysis 8-7-09
 * Dot Blot Results 8-7-09
 * Biotinylation of ab after removal of sodium azide 8-12-09
 * dot blot to check biotinylation 8-13-09
 * dot blot w/hrp 8-13-09
 * dot blot imaging 8-15-09
 * dot blot procedure plan 8-13-09
 * results of m13cp gel 8-18-09
 * phage ELISA 8-19-09 to 8-20-09
 * m13 plasmid glycerol stock 8-21-09
 * phage ELISA Plan 8-21-09
 * Dilution series of alexafluor 8-21-09
 * Phage ELISA Results 8-24-09 to 8-25-09
 * phage ELISA Results 8-25-09
 * biotinylation of phage ab 8-28-09
 * Phage ELISA 9-1-09
 * Phage ELISA Results 9-1-09
 * Estimating Phage Concentration from absorbance 9-3-09
 * m13cp verification 9-3-09
 * titration of scFv 9 9-3-09
 * Results of scFv9 Titration 9-6-09
 * Phage on Array schematic 9-7-09
 * phage on array calculations 9-7-09
 * phage array protocol 9-17-09
 * naive library plasmid prep 9-17-09
 * preparing to make electrically competent m13cp 9-22-09
 * phage mice immunization 9-23-09
 * preparation of electrocompetent e coli protocol 9-24-09
 * preparation of electrocompetent m13cp and m13cp-CT 9-24-09
 * transformation of naive library into helper strains 9-25-09
 * retransformed library into helper strains 9-26-09
 * check on naive lib into helper cell transformation 9-28-09
 * glycerol stock to phage protocol 9-29-09
 * glycerol stock of phage 9-29-09
 * glycerol stock to phage 10-1-09
 * titration of 6 monoclonal phages 10-5-09
 * result of 6 monoclonal phage titration 10-6-09
 * 2nd phage mice immunization 10-7-09
 * m13cp verification 2nd attempt 10-7-09
 * plasmid extraction w/spin minikit 10-8-09
 * plasmid extraction w/qiagen kit 10-9-09
 * sequencing monoclonal phage 10-9-09
 * restriction digest of helper plasmids 10-9-09
 * planning naive library amplification 10-12-09
 * naive library amplification plan 10-13-09
 * mouse cheekbleed/naive library amplification 10-14-09
 * monoclonal phage sequence analysis/naive library amplification 10-15-09
 * naive library titration result 10-17-09
 * naive library titration 10-19-09
 * anti-M13 Mouse sera ELISA w/HRP 10-19-09
 * anti-M13 Mouse sera results 10-19-09
 * cancer B cell isolation 10-21-09
 * mouse sera phage ELISA preparation 10-21-09
 * mouse sera phage ELISA 10-21-09
 * mouse sera phage ELISA results 10-21-09
 * 3rd phage mouse immunization 10-23-09
 * naive library verification 10-22-09
 * cancer b cell rna extraction 10-22-09
 * helper and PCANTAB gel 10-26-09
 * anti-m13 mouse sera ELISA 10-29-09
 * mouse sera ELISA results 10-30-09
 * pcr planning 11-2-09
 * anti-m13 mouse euthanization 11-4-09
 * cr of m13cp-CT-2 sample 11-6-09
 * result of m13cp-CT-2 PCR 11-9-09
 * pcr of m13cp-1 and m13cp-2 11-9-09
 * gel extraction of m13cp-1 11-12-09
 * phage insert check pcr results 11-13-09
 * ELISA for mouse sera titer 11-13-09
 * ELISA Mouse sera titer results 11-13-09
 * rna extraction schematic 3-8-10


 * Tumor and B cell RNA Extraction 6-14-10
 * RNA Isolation from B cells of Naive Mouse 6-14-10
 * Tumor, spleen, and blood collection 6-18-10
 * Isolation of B cells 6-18-10
 * Isolation of RNA 6-21-10
 * Naive B RNA Check 6-22-10
 * RNA Isolation from Tumor Tissue 6-24-10
 * Rerun of tumor RNA Gel 6-25-10
 * Preparation of cDNA from B cell RNA 7-1-10
 * Beta actin check of cDNA 7-2-10
 * Amplification of Antibody Gene Regions from cDNA 7-6-10 to 7-7-10
 * PCR of Antibody Gene Region 7-8-10
 * cDNA Synthesis 7-8-10
 * Naive Spleen Processing 7-19-10
 * RNA from Naive B cells 7-19-10
 * cDNA from Naive B RNA 7-20-10
 * PCR to Amplify antibody gene regions 7-21-10 to 7-22-10
 * Gel of Antibody Genes 7-23-10
 * Rerun of PCR to amplify ab genes 7-26-10 to 7-26-10
 * test PCR of ab genes 7-29-10
 * gel check of ab gene PCR 7-30-10
 * RNA Stability Test 7-30-10
 * PCR of Tumor ab genes 8-2-10
 * test of pcr with Hojoon's cDNA 8-3-10
 * pcr of ab genes with lower primer conc. 8-4-10
 * pcr of ab genes with more template 8-5-10
 * process spleens of smc1 mice 8-9-10
 * optimization of pcr of ab genes 8-10-10
 * optimization of pcr of ab genes 8-10-10
 * amplifying with all 68 primers for naive 8-10-10
 * pcr ab genes (tumor, smc1, 6-21) 8-17-10 to 8-21-10
 * pcr ab genes (tumor, smc1, 6-21) with extra primer and polymerase to amplify faint bands
 * pcr ab genes gel images 8-21-11
 * extra thermocycles for rest of pcr plates 8-23-10
 * summary of pcrs 8-23-10
 * cDNA synthesis of smc1 RNA 8-24-10
 * amplification of smc1 ab genes 8-25-10
 * additional pcr cycles for troublesome primer pairs 8-27-10
 * pcr to amplify genes with difficult primers 8-27-10
 * gel extraction of ab genes 8-30-10
 * pcr purification of ab genes 8-30-10
 * pcr purification of ab genes 8-31-10
 * conclusion of analysis of ab genes 9-8-10
 * addition of linker to 1-78 9-9-10
 * addition of linker to 1-78 9-9-10
 * gel extraction of ab genes w/linker 9-10-10
 * overlapping pcr 9-10-10
 * flanking primers for overlapping pcr 9-11-10
 * check overlapping pcr and amplification of tumor ab genes 9-12-10
 * pcr to add linker to 1-78 genes 9-13-10
 * gel extraction of ab genes with linker 9-14-10
 * DH10B competent cell prep 9-15-10
 * Amplification of linked heavy and light 9-15-10
 * check of linked heavy and light 1-78 ab genes 9-16-10
 * check of tumor ab genes 9-16-10
 * pcr purification of tumor ab genes 9-20-10
 * check of transformation efficiency 9-20-10
 * gel extraction of tumor ab genes 9-21-10
 * naive ab gene amplification and first 10 tumor ab gene amplification 9-21-10
 * additional thermocycles for naive and tumor genes 9-22-10
 * gel of naive ab genes and 1st 10 tumor ab genes 9-23-10
 * tumor ab genes 7-10 9-23-10
 * DH10B competent cell preparation 9-24-10
 * gel extraction and pcr purification 9-29-10
 * nanodrop concentrations of naive 9-29-10
 * pcr for last few tumor ab genes 10-1-10
 * pcr purification and gel extraction 10-2-10
 * pool of primers w/SMC1 DNA 10-4-10
 * pcr of gel extracted ab smc1 gene 10-6-10
 * Construction of tumor scFv DNA 10-7-10 to 10-8-10
 * Amplification of Tumor scFV DNA 10-10-10
 * Construction of naive, SMC1 and 1-78 scFV DNA 10-11-10
 * Link heavy and light chains of 1-78 Naive and SMC1 DNA 10-13-10
 * Amplification of linked heavy and light chain genes 10-14-10
 * Amplification of scFV DNA 10-15-10
 * Restriction of 1-78 scFV with sfiI 10-18-10
 * Better picture of scFV DNA 10-20-10
 * Final scFV DNA gel 10-22-10
 * Final scFV DNA gel with fresh buffer 10-25-10
 * Gel extraction of antibody genes with linker 10-27-10
 * Construction of scFV 10-29-10
 * Gel extraction of final scFV 10-30-10
 * Amplified linked scFV fragments 11-2-10
 * scFV construction 11-4-10
 * Tumor lambda construction 11-5-10 to 11-8-10
 * Gel of final scFV DNA 11-11-10
 * Restriction digest of PCANTAB5E 11-15-10 to 11-16-10
 * Isolation of scFv DNA 11-18-10
 * Investigation of PCANTAB5E 11-17-10
 * Ligate PCANTAB with rCompTT 4-8-11
 * Ethanol precipitate ligation 4-11-11
 * Preparation of phage helper competent cells 4-19-11 to 4-21-11
 * Transformation of helper competent cells 4-22-11
 * Preparation to make competent cells 4-26-11
 * Preparation of Electrocompetent ER2738 w/M13cp-CT 4-29-11
 * Transformation efficiency and glycerol stock 5-4-11
 * Colony PCR for F1 ori 5-24-11
 * Helper ER2738 W/M13cp-CT preparation 7-20-11
 * Transformation of ER2738 M13cp-CT 7-26-11
 * Electrocompetent ER2738 M13cp-CT preparation 8-2-11
 * Test transformation "fast freeze H2O" 8-3-11
 * Electrocompetent cell preparation 8-4-11
 * Electrocomporation of ER2738 8-5-11
 * Transformation results 8-6-11
 * Test ligation for ER2738 8-11-11
 * EtOH precipitation and transformation 8-12-11
 * Cut PCANTAB5E 8-16-11
 * Recut PCANTAB for commercially competent cells 8-18-11
 * Gel extraction and ligation 8-19-11
 * Ethnal precipitation and transformation 8-20-11
 * Sfi I mouse SMC1fs 8-25-11
 * Restriction digest table 8-25-11
 * PCR purification/Gel extraction 8-25-11
 * Ethanol precipitation 8-26-11
 * Eletroporation of mouse SMC1fs scFV 8-26-11
 * Minipreps SMC1fs scFV 8-28-11
 * Colony PCR of SMC1fs scFV 8-29-11
 * Colony PCR test 9-1-11
 * Advantage PCR of miniprep 9-1-11
 * Cut SMC1fs scFV PCANTAB plasmids 9-6-11
 * BamHI and Hind III double digest 9-7-11
 * Restriction Digest and overnight Gel 9-7-11
 * Not I restriction digest check 9-10-11
 * Hind III & EcoRI digest 9-9-11
 * Not I digest with both buffers 9-9-11
 * Basic field inversion electrophoresis test 9-10-11
 * Cut PCANTAB for stuffer (SacI and PstI) 9-15-11
 * Cut PCANTAB with SacI and NotI 9-19-11
 * pGEX-SMC1fs cut with NotI 9-22-11
 * pRSET cut with SacI and NotI 9-24-11
 * Gel of pRSET sequential cut 9-24-11
 * Ligation 9-24-11
 * Ethanol precipitation PCANTAB pRSET 9-26-11
 * Electroporation protocol 9-26-11
 * tRNA- carrier preparation 10-3-11
 * PCANTAB -PRSET digest 10-4-11
 * Potential PCANTAB pRSET 10-5-11
 * Ethanol precipitation of PCANTAB maxiprep 10-19-11
 * Ethanol precipitation PCANTAB pRSET 9-26-11
 * Electroporation protocol 9-26-11
 * tRNA- carrier preparation 10-3-11
 * PCANTAB -PRSET digest 10-4-11
 * Potential PCANTAB pRSET 10-5-11
 * Ethanol precipitation of PCANTAB maxiprep 10-19-11


 * Check of PCANTAB Maxiprep 11-1-11
 * PTAB Fragments into pJET 11-3-11
 * PCR of PCANTAB pJET clones 11-7-11
 * Gel and Miniprep 11-8-11


 * Cut Vector with SpeI and NotI 061412
 * Modification of pComb3xSS vector 6-25-12
 * Diagram of nucleotide changes 6-26-12
 * Cut pComb with SfiI and NotI 7-6-12
 * PCR of pComb fragment 7-7-12
 * pComb In-Fusion 7-9-12
 * Resequence pComb NotI Modification 7-19-12
 * Miniprep pComb Cultures 7-21-12
 * added polyT near lacI in pComb 8-27-12
 * pcomb mod with 2nd round primers 8-29-12
 * modified pcomb electroporation 8-31-12
 * mpComb Sequencing 9-5-12
 * recut MN pComb 9-18-12 (MN is for modified NotI)
 * pComb fragment PCR round 1 9-19-12
 * pComb fragment PCR round 2 9-20-12
 * pComb In-Fusion 9-21-12
 * Extracted mpComb Plasmids 9-23-12
 * Cut pComb and rCompTT 10-4-12
 * NotI Digestion 10-5-12
 * ligate practice fragment 10-9-12
 * test for pComb Insert 10-15-12
 * pComb insert gel 10-15-12
 * pComb digest 10-16-12
 * CIAP treatment 10-16-12
 * pComb Gel extraction 10-16-12
 * pComb ligation 10-19-12
 * transformation of pcomb ligation 10-24-12
 * pcomb ligation and transformation 10-25-12
 * verify rcompTT ligation 10-29-12
 * rCompTT Ligation verification 10-29-12
 * sequence new pComb plasmid 10-31-12
 * pComb digest 10-16-12
 * CIAP Treatment 1-16-12
 * pComb Gel Extraction 10-16-12
 * pComb Ligation 10-19-12
 * Transformation of pComb Ligation 10-24-12
 * pComb Ligation and transformatio 10-25-12
 * verify rCompTT Ligation 10-29-12
 * rCompTT ligation verification 10-29-12
 * sequence new pcomb plasmid 10-31-12
 * pComb MN linearization 11-13-12
 * Construction of fragment for pComb 11-16-12
 * pComb MN linearization 11-16-12
 * Stellar electroporation 11-20-12
 * pComb linearization check 11-20-12
 * linearization with more polymerase 11-21-12
 * check of linearization 11-22-12
 * polymerase system test 11-26-12
 * check polymerase test 11-27-12
 * pComb linearization check 11-27-12
 * pComb linearization with PrimeSTAR 11-28-12
 * In-Fusion with linearized pComb 12-4-12
 * pComb relinearization 12-13-12
 * pComb linearization 1-14-13
 * In-fusion of linearized pComb 1-15-13
 * linearized pcomb gel extraction 1-16-13
 * sequencing pcomb candidates 1-18-13
 * sequenced pComb 1-29-13

Note there seems to be some dates missing here such as "Cut rCompTT w/SfiI 2-24-11)