Difference between revisions of "Team:Aachen/Notebook/Documentation/Methanol Monocistronic Diversity Library"
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! gene !! RDP parts !! product in !! expected length / result !! plasmid ID !! cryo ID | ! gene !! RDP parts !! product in !! expected length / result !! plasmid ID !! cryo ID | ||
|- | |- | ||
− | | Mdh || (mdh.TER.AP00) #XOXM# + #PRDW# + #PLTB# + #PQT6# + #PVFL# || #K3YV# ; empty|| 3497 bp / | + | | Mdh || (mdh.TER.AP00) #XOXM# + #PRDW# + #PLTB# + #PQT6# + #PVFL# || #K3YV# ; empty|| 3497 bp / no evaluable result|| #CDPA#, #ZYBC# || #SF63#, #MBRR# |
|- | |- | ||
− | | Mdh || (mdh.TER.AP04) #XOXM# + #PRDW# + #PLTB# + #WPW6# + #PVFL# || #AVQ6#; empty || 3497 bp / | + | | Mdh || (mdh.TER.AP04) #XOXM# + #PRDW# + #PLTB# + #WPW6# + #PVFL# || #AVQ6#; empty || 3497 bp / no evaluable result || #YNOB#, #1KSW# || #DCN9#, #WHBF# |
|- | |- | ||
− | | Mdh || (mdh.TER.AP10) #XOXM# + #PRDW# + #PLTB# + #3S8V# + #PVFL# || #ZE1A#; empty || 3497 bp / | + | | Mdh || (mdh.TER.AP10) #XOXM# + #PRDW# + #PLTB# + #3S8V# + #PVFL# || #ZE1A#; empty || 3497 bp / no evaluable result|| #EBWO#, #936C# || #8KAA#, #EB88# |
|- | |- | ||
− | | Mdh || (mdh.TER.AP19) #XOXM# + #PRDW# + #PLTB# + #MBWS# + #PVFL# || #NDKS#; empty || 3497 bp / | + | | Mdh || (mdh.TER.AP19) #XOXM# + #PRDW# + #PLTB# + #MBWS# + #PVFL# || #NDKS#; empty || 3497 bp / no evaluable result || #1MC6#, #ESQD# || #48C8#, #8BZ9# |
|} | |} | ||
Line 22: | Line 22: | ||
==15-08-09== | ==15-08-09== | ||
− | '''Gel control of | + | '''Gel control of mdh precursor plasmids''': |
− | *no good picture could be taken because the UV light destroyed the DNA; picture of it will be taken tomorrow | + | *no good picture could be taken because the UV light destroyed the DNA; picture of it will be taken tomorrow |
− | + | *plasmid purification and cryo cultures of mdh precursor plasmids | |
− | *plasmid | + | |
==15-08-10== | ==15-08-10== | ||
− | *make | + | *make cryo cultures and purify plasmids of mdh precursor plasmids |
− | *Measure concentrations of | + | *Measure concentrations of purified mdh precursor plasmids |
− | *Gel control of | + | *Gel control of mdh precursor plasmids |
− | + | *Pipett sequencing of mdh precursor plasmids | |
− | *Pipett sequencing of | + | |
− | * test digest of | + | * test digest of mdh precursor plasmids: |
{|class="wikitable" | {|class="wikitable" | ||
Line 55: | Line 53: | ||
| H || #ESQD# || 2226 / 1279 || 2226 / 1279 | | H || #ESQD# || 2226 / 1279 || 2226 / 1279 | ||
|- | |- | ||
− | | | + | |} |
+ | |||
+ | digests stored in falcon #PSHF# | ||
+ | |||
+ | ==15-08-11== | ||
+ | * sequencing of following samples: #1MC6#, #ESQD#, | ||
+ | * '''oligo assembly''': BBa_B1002-RDP (#HTSR#), BBa_B1006-RDP (#8AMS#), AP19-RDP (#MBWS#) | ||
+ | **heated on 90 °C, then linear cool down in 45 minutes to room temperature | ||
+ | |||
+ | '''assembly of hps Precursors''' | ||
+ | {|class="wikitable" | ||
+ | ! gene !! RDP parts !! product in | ||
|- | |- | ||
− | | | + | | hps || (hps.B1006.AP00) #RYPQ# + #VO4C# + #8AMS# + #M1ZY# + #OZD1# || #V9H9# |
|- | |- | ||
− | | | + | | hps || (hps.B1006.AP04) #RYPQ# + #VO4C# + #8AMS# + #HB96# + #OZD1# || #XKX6# |
|- | |- | ||
− | | | + | | hps || (hps.B1006.AP10) #RYPQ# + #VO4C# + #8AMS# + #4T9A# + #OZD1# || #SNDK# |
|- | |- | ||
− | | | + | | hps || (hps.B1006.AP19) #RYPQ# + #VO4C# + #8AMS# + #XYD9# + #OZD1# || #BC4V# |
+ | |} | ||
+ | |||
+ | |||
+ | *heat shock transformation in DH5alpha with 5 µl of DNA | ||
+ | |||
+ | ==15-08-12== | ||
+ | *there were colonies on the transformation plates of the hps precursors | ||
+ | *Oligos from 15-08-11: put them on 2% agarose: | ||
+ | (good results: 0,8µl DNA and more than 3,5 µl 50bp Ladder) | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! part !! expected lenght !! result | ||
|- | |- | ||
− | | | + | | BBa_B1002-RDP (#HTSR#) || 45 bp || fragments of ca. 50bp |
+ | |- | ||
+ | | BBa_B1006-RDP (#8AMS#) || 50 bp || fragments of ca. 50bp | ||
+ | |- | ||
+ | | AP19-RDP(#MBWS#) || 47 bp || fragments of ca. 50bp | ||
|} | |} | ||
− | + | ===Assembly of phi precursors=== | |
+ | |||
+ | {|class="wikitable" | ||
+ | ! gene !! RDP parts !! product in | ||
+ | |- | ||
+ | | phi || (phi.B1002.AP00) || #VVSC# | ||
+ | |- | ||
+ | | phi || (phi.B1002.AP04) || #BNHW# | ||
+ | |- | ||
+ | | phi || (phi.B1002.AP10) || #D1LL# | ||
+ | |- | ||
+ | | phi || (phi.B1002.AP19) || #EB8T# | ||
+ | |} | ||
+ | *each 5µl transformed in DH5alpha | ||
+ | *masterplates and colony PCR will follow | ||
+ | |||
+ | ===colony PCR of mdh precursor clones=== | ||
+ | * primers: #MSP2# and #94LB# | ||
+ | * elongation time: 1'00'' | ||
+ | * annealing temperature: 58°C | ||
+ | * expected length: 920 bp | ||
+ | {|class="wikitable" | ||
+ | ! ID !! description !! positive clones | ||
+ | |- | ||
+ | | A 1-12 || mdh.Ter.AP00 || no | ||
+ | |- | ||
+ | | B 1-15 || mdh.Ter.AP04 || 5 | ||
+ | |- | ||
+ | | C 1-12 || mdh.Ter.AP10 || no | ||
+ | |- | ||
+ | | D 1-11 || mdh.Ter.AP19 || 2, 4 | ||
+ | |} | ||
+ | |||
+ | ==15-08-13== | ||
+ | ===tasks:=== | ||
+ | * colony PCR of hps precursors | ||
+ | * colony PCR of mdh precursors (again) | ||
+ | * masterplates of phi precursors | ||
+ | * overnight cultures of positive colony PCR clones | ||
+ | |||
+ | ====Colony PCRs==== | ||
+ | |||
+ | '''mdh precursor colony PCR:''' | ||
+ | * Primers: #MSP2#, #94LB# | ||
+ | {|class="wikitable" | ||
+ | ! step !! time !! Temp [°C] | ||
+ | |- | ||
+ | | denature || 5'00'' || 94 | ||
+ | |- | ||
+ | | denature || 0'30'' || 94 | ||
+ | |- | ||
+ | | anneal || 0'30'' || 55 | ||
+ | |- | ||
+ | | elongate || 1'00'' || 68 | ||
+ | |- | ||
+ | | elongate || 2'00'' || 68 | ||
+ | |- | ||
+ | | store || forever || 16 | ||
+ | |} | ||
+ | '''hps precursor colony PCR:''' | ||
+ | * Primers: #HSP1#, #94LB# | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! step !! time !! Temp [°C] | ||
+ | |- | ||
+ | | denature || 5'00'' || 94 | ||
+ | |- | ||
+ | | denature || 0'30'' || 94 | ||
+ | |- | ||
+ | | anneal || 0'30'' || 55 | ||
+ | |- | ||
+ | | elongate || 0'40'' || 68 | ||
+ | |- | ||
+ | | elongate || 2'00'' || 68 | ||
+ | |- | ||
+ | | store || forever || 16 | ||
+ | |} | ||
+ | |||
+ | '''Results:''' | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! construct ID !! construct description !! expected length [bp] !! positive clones | ||
+ | |- | ||
+ | | A 1-8 || mdh AP00 precursor || 920 bp || (A16), A18 | ||
+ | |- | ||
+ | | B 1-8 || mdh AP04 precursor || 920 bp || B15, (B17), (B18) | ||
+ | |- | ||
+ | | C 1-8 || mdh AP10 precursor || 920 bp || - | ||
+ | |- | ||
+ | | D 1-8 || mdh AP19 precursor || 920 bp || D12, D14 | ||
+ | |- | ||
+ | | E 1-8 || hps AP00 precursor || 579 bp || - | ||
+ | |- | ||
+ | | F 1-8 || hps AP04 precursor || 579 bp || - | ||
+ | |- | ||
+ | | G 1-8 || hps AP10 precursor || 579 bp || - | ||
+ | |- | ||
+ | | H 1-8 || hps AP19 precursor || 579 bp || - | ||
+ | |} | ||
+ | |||
+ | ==15-08-14== | ||
+ | ===tasks:=== | ||
+ | * colony PCR of all phi precursors | ||
+ | * colony PCR of all hps precursors | ||
+ | * colony PCR of mdh AP10 precursor | ||
+ | * purify plasmid of: | ||
+ | ** mdh AP00 precursors (A16, A18) | ||
+ | ** mdh AP04 precursors (B15, B17, B18) | ||
+ | ** mdh AP19 precursors (D12, D14) | ||
+ | * label gels of yesterday | ||
+ | * BsaI digest clean-up | ||
+ | |||
+ | ===purified plasmids and cryo cultures of mdh precursors=== | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! construct !! Cryo ID !! Plasmid ID !! Plasmid conc. [ng/µl] | ||
+ | |- | ||
+ | | mdh AP00 #16 || #Q6XM# || #CZ83# || 125,5 | ||
+ | |- | ||
+ | | mdh AP00 #18 || #6F1N# || #XNTZ#|| 171,5 | ||
+ | |- | ||
+ | | mdh AP04 #15 || #848Y# || #SRYN#|| 338 | ||
+ | |- | ||
+ | | mdh AP04 #17 || #FCKK# || #TK9X#|| 134 | ||
+ | |- | ||
+ | | mdh AP04 #18 || #3C1N# || #C8EB#|| 147,5 | ||
+ | |- | ||
+ | | mdh AP19 #12|| #XYM1# || #VMBV#|| 72 | ||
+ | |- | ||
+ | | mdh AP19 #14 || #RB99# || #4P6A#|| 101,5 | ||
+ | |} | ||
+ | |||
+ | ====Colony PCRs==== | ||
+ | '''Program:''' | ||
+ | {|class="wikitable" | ||
+ | ! step !! time !! T [°C] | ||
+ | |- | ||
+ | | denature || 5'00'' || 94 | ||
+ | |- | ||
+ | | denature || 0'30'' || 94 | ||
+ | |- | ||
+ | | annealing || 0'30'' || 55 | ||
+ | |- | ||
+ | | elongation || varying || 68 | ||
+ | |- | ||
+ | | final elongation || 2'00'' || 68 | ||
+ | |- | ||
+ | | store || forever || 16 | ||
+ | |} | ||
+ | |||
+ | '''Results table:''' | ||
+ | {|class="wikitable" | ||
+ | ! construct !! Primers !! elongation time !! expected length [bp] !! tested clones !! positive clones | ||
+ | |- | ||
+ | | mdh.AP10 precursor || #MSP2#, #94LB# || 1'00'' || 920 || 1, 2, 3, 4, 5, 19, 20, 21 || - | ||
+ | |- | ||
+ | | hps.AP00 precursor || #W6WP#, #94LB# || 1'13'' || 1160 || 9-16 || 15 | ||
+ | |- | ||
+ | | hps.AP04 precursor || #W6WP#, #94LB# || 1'13'' || 1160 || 9-16 || 9 | ||
+ | |- | ||
+ | | hps.AP10 precursor || #W6WP#, #94LB# || 1'13'' || 1160 || 9-14 || 11, 13, 14 | ||
+ | |- | ||
+ | | hps.AP19 precursor || #W6WP#, #94LB# || 1'13'' || 1160 || 9-16 || 9, 13, 14 , 15, 16 | ||
+ | |- | ||
+ | | phi.AP00 precursor || #W6WP#, #94LB# || 1'13'' || 1074 || 1-8 || - | ||
+ | |- | ||
+ | | phi.AP04 precursor || #W6WP#, #94LB# || 1'13'' || 1074 || 1-8 || - | ||
+ | |- | ||
+ | | phi.AP10 precursor || #W6WP#, #94LB# || 1'13'' || 1074 || 1-8 || 2, 3, 4, 10 | ||
+ | |- | ||
+ | | phi.AP19 precursor || #W6WP#, #94LB# || 1'13'' || 1074 || 1-6 || 5 | ||
+ | |} | ||
+ | |||
+ | ====Assembly of MDH precursors==== | ||
+ | * this time with blockers; for higher coupling efficiency | ||
+ | **Assembly product mdh.AP00: #N1O1# | ||
+ | **Assembly product mdh.AP04: #VYM6# | ||
+ | **Assembly product mdh.AP10: #Q9QQ# | ||
+ | **Assembly product mdh.AP19: #ONWS# | ||
+ | |||
+ | ==15-08-15== | ||
+ | ====Tasks:==== | ||
+ | * purify plasmid of prouven clones from yesterday | ||
+ | * '''make masterplates of MDH precursors (new assembly, this time with blockers)''' | ||
+ | * assembly of hps and phi precursors (with blockers) | ||
+ | * tidy up fridge | ||
+ | * do transformation of hps and phi precursors | ||
+ | * do transformation of mdh.AP04 and mdh.AP10 precursors | ||
+ | |||
+ | ====Plasmid purification of hps and phi precursors==== | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! construct !! Cryo ID !! Plasmid ID !! Plasmid conc. [ng/µl] | ||
+ | |- | ||
+ | | hps AP19 #14 || #FOF4# || #B3PQ# || 170 | ||
+ | |- | ||
+ | | hps AP10 #14 || #638V# || #P4SB#|| 148 | ||
+ | |- | ||
+ | | hps AP10 #11 || #4ZSB# || #P89F#|| 143,5 | ||
+ | |- | ||
+ | | hps AP04 #9 || #NY1C# || #1TMT#|| 190,5 | ||
+ | |- | ||
+ | | hps AP19 #15 || #SM3L# || #NFOS#|| 204 | ||
+ | |- | ||
+ | | hps AP00 #15|| #HAKR# || #6ACO#|| 126,5 | ||
+ | |- | ||
+ | | hps AP19 #13 || #COMY# || #VSWC#|| 261 | ||
+ | |- | ||
+ | | hps AP19 #9 || #OZ4O# || #L8Y4#|| 171,5 | ||
+ | |- | ||
+ | | hps AP10 #13 || #8P4S# || #4QLE#|| 238,5 | ||
+ | |- | ||
+ | | hps AP19 #16|| #CSLZ# || #6P68#|| 121,5 | ||
+ | |- | ||
+ | | phi AP10 #3 || #BV3E# || #L9E9#|| 161 | ||
+ | |- | ||
+ | | phi AP10 #4 || #OFHN# || #FVVW#|| 171,5 | ||
+ | |- | ||
+ | | phi AP19 #5 || #E6HX# || #HA1W#|| 192,5 | ||
+ | |} | ||
+ | |||
+ | ===Assembly of hps precursors=== | ||
+ | * this time with blockers (better coupling efficiency) | ||
+ | **Assembly product hps.AP00: #WQPR# | ||
+ | **Assembly product hps.AP04: #6WOE# | ||
+ | **Assembly product hps.AP10: #XHXQ# | ||
+ | **Assembly product hps.AP19: #VM1V# | ||
+ | |||
+ | ===Assembly of phi precursors=== | ||
+ | * this time with blockers (better coupling efficiency) | ||
+ | **Assembly product phi.AP00: #AFA4# | ||
+ | **Assembly product phi.AP04: #8FXO# | ||
+ | **Assembly product phi.AP10: #KCDE# | ||
+ | **Assembly product phi.AP19: #QHM6# | ||
+ | |||
+ | ====Transformations==== | ||
+ | {|class="wikitable" | ||
+ | ! # !! description !! assembly product | ||
+ | |- | ||
+ | | 1 || hps.AP00 precursor || #WQPR# | ||
+ | |- | ||
+ | | 2 || hps.AP04 precursor || #6WOE# | ||
+ | |- | ||
+ | | 3 || hps.AP10 precursor || #XHXQ# | ||
+ | |- | ||
+ | | 4 || hps.AP19 precursor || #VM1V# | ||
+ | |- | ||
+ | | 5 || phi.AP00 precursor || #AFA4# | ||
+ | |- | ||
+ | | 6 || phi.AP04 precursor || #8FXO# | ||
+ | |- | ||
+ | | 7 || phi.AP10 precursor || #KCDE# | ||
+ | |- | ||
+ | | 8 || phi.AP19 precursor || #QHM6# | ||
+ | |- | ||
+ | | 9 || mdh.AP04 precursor || #VYM6# | ||
+ | |- | ||
+ | | 10 || mdh.AP10 precursor || #Q9QQ# | ||
+ | |} | ||
+ | |||
+ | ==15-08-16== | ||
+ | |||
+ | * make masterplates of yesterdays transformations | ||
+ | |||
+ | ==15-08-17== | ||
+ | ===Precursor plasmids:=== | ||
+ | {|class="wikitable" | ||
+ | ! construct !! cryo IDs !! plasmid IDs !! screening result | ||
+ | |- | ||
+ | | mdh00 precursor || #L99Q#, #VLO3#, #EHQC#, #OD19#, #FHEL#, #MA9W# || #HR4E#, #ZZEE#, #3HPD#, #M1VB#, #QDVN#, #AHRM# || results on table from tomorrow | ||
+ | |- | ||
+ | | mdh04 precursor || #OB4C#, #TVND#, #1S1V#, #COXQ#, #3NZQ#, #YZXD# || #NEKS#, #OX44#, #P9HA#, #9CBA#, #SYMS#, #YY6O# || | ||
+ | |- | ||
+ | | mdh19 precursor || #HBWZ#, #K3V6#, #CYF6#, #S1HT#, #TKO4#, #4MS8# || #BY9L#, #DNRY#, #KV18#, #1KEA#, #SPHL#, #N1BM# || | ||
+ | |- | ||
+ | | hps00 precursor || #AVVO#, #X9PR#, #M4H8#, #44WP#, #DE4A#, #QMRB# || #XVYK#, #CMC1#, #ERZK#, #WXO8#, #D8PY#, #8AFD# || | ||
+ | |- | ||
+ | | hps04 precursor || #8SW4#, #RSH9#, #1C8L#, #E1OQ#, #Y3TR#, #3BLV# || #YE8A#, #DERR#, #DVRP#, #FKNX#, #N3E4#, #HNV4# || | ||
+ | |- | ||
+ | | hps10 precursor || #R8YT#, #QRZW#, #KVYA#, #NEFO#, #QN64#, #NSSM# || #1EQ3#, #PA13#, #FAES#, #Z64V#, #WSAD# || | ||
+ | |- | ||
+ | | phi00 precursor || #TNBW#, #VSQN#, #YPFO#, #EELW#, #9H4T#, #VLHL# || #D6QY#, #931O#, #QWQP#, #AERW#, #ORX1#, #ZCTY# || | ||
+ | |- | ||
+ | | phi04 precursor || #4MNP#, #DE96#, #YKZT#, #AQF8#, #ETTS#, #8COE# || #FXSC#, #P1O1#, #8CZD#, #EZYZ#, #VPP4#, #3EZP# || | ||
+ | |- | ||
+ | | phi10 precursor || #SQ9N#, #AO6C#, #9RZL#, #48P3#, #PPWW#, #DB3R# || #FTX4#, #VM6Z#, #V4CA#, #XW1Y#, #AZND#, #WCF3# || | ||
+ | |- | ||
+ | | phi19 precursor || #3WKA#, #94OY#, #TVM1#, #KPCQ#, #FB9L#, #HWM4# || #THQA#, #BND4#, #6H3Z#, #ME9O#, #39CF#, #RABS# || | ||
+ | |} | ||
+ | |||
+ | ===further plasmids:=== | ||
+ | * HPS19 precursor | ||
+ | ** cryo: #ABCE# | ||
+ | ** plasmid: #TTC4# | ||
+ | |||
+ | ==15-08-18== | ||
+ | ===Morning=== | ||
+ | Plasmid purifications and sequencing. | ||
+ | Pipetting calculation table is ready at: '''\WetLab\Documentation\Methanol\15-08-18 Sequencings\''' | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! construct & clone number !! cryo ID !! plasmid ID !! sequencing sample numbers !! sequencing result !! further procedure | ||
+ | |- | ||
+ | | mdh.TER.AP00 in pSB1CRDP #1 || #L99Q# || #HR4E# || 15 || MUTATION || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP00 in pSB1CRDP #2 || #OD19# || #M1VB# || 16 || MUTATION || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP00 in pSB1CRDP #6 || #MA9W# || #AHRM# || 17 || MUTATION || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP04 in pSB1CRDP #1 || #OB4C# || #NEKS# || 18 || SPACER MUTATION || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP04 in pSB1CRDP #2 || #TVND# || #OX44# || 19 || VALID || Keep | ||
+ | |- | ||
+ | | mdh.TER.AP04 in pSB1CRDP #3 || #1S1V# || #P9HA# || 20 || VALID || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP19 in pSB1CRDP #1 || #HBWZ# || #BY9L# || 21 || VALID || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP19 in pSB1CRDP #2 || #K3V6# || #DNRY# || 22 || VALID || Discard | ||
+ | |- | ||
+ | | mdh.TER.AP19 in pSB1CRDP #3 || #CYF6# || #KV18# || 23 || VALID || Keep | ||
+ | |- | ||
+ | | hps.B1006.AP00 in pSB1CRDP #3 || #M4H8# || #ERZK# || 24 || VALID || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP00 in pSB1CRDP #4 || #44WP# || #WXO8# || 25 || VALID || Keep | ||
+ | |- | ||
+ | | hps.B1006.AP00 in pSB1CRDP #5 || #DE4A# || #D8PY# || 26 || NO CONTIGS FOUND || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP04 in pSB1CRDP #2 || #RSH9# || #DERR# || 27 || MUTATION || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP04 in pSB1CRDP #3 || #1C8L# || #DVRP# || 28 || MUTATION || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP04 in pSB1CRDP #6 || #3BLV# || #HNV4# || 29 || MUTATION || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP10 in pSB1CRDP #2 || #QRZW# || #1EQ3# || 30 || NO CONTIGS FOUND || Discard | ||
+ | |- | ||
+ | | hps.B1006.AP10 in pSB1CRDP #4 || #NEFO# || #FAES# || 31 || NO CONTIGS FOUND|| Discard | ||
+ | |- | ||
+ | | hps.B1006.AP19 in pSB1CRDP #1 || #ABCE# || #TTC4# || 32 || MUTATION || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP00 in pSB1CRDP #2 || #VSQN# || #931O# || 33 || VALID || Keep | ||
+ | |- | ||
+ | | phi.B1002.AP00 in pSB1CRDP #3 || #YPFO# || #QWQP# || 34 || NO CONTIGS FOUND || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP00 in pSB1CRDP #4 || #EELW# || #AERW# || 35 || MUTATION || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP04 in pSB1CRDP #1 || #4MNP# || #FXSC# || 36 || NO CONTIGS FOUND || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP04 in pSB1CRDP #3 || #YKZT# || #8CZD# || 37 || PROMOTER OK || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP04 in pSB1CRDP #4 || #AQF8# || #EZYZ# || 38 || VALID || Keep | ||
+ | |- | ||
+ | | phi.B1002.AP10 in pSB1CRDP #3 || #9RZL# || #V4CA# || 39 || VALID || Keep | ||
+ | |- | ||
+ | | phi.B1002.AP10 in pSB1CRDP #4 || #48P3# || #XW1Y# || 40 || VALID || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP10 in pSB1CRDP #6 || #DB3R# || #WCF3# || 41 || VALID || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP19 in pSB1CRDP #2 || #94OY# || #BND4# || 42 || NO CONTIGS FOUND || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP19 in pSB1CRDP #5 || #FB9L# || #39CF# || 43 || MUTATION || Discard | ||
+ | |- | ||
+ | | phi.B1002.AP19 in pSB1CRDP #6 || #HWM4# || #RABS# || 44 || VALID || Keep | ||
+ | |} | ||
+ | |||
+ | |||
+ | ==15-08-20== | ||
+ | ===missing Precursors:=== | ||
+ | |||
+ | do cryo and plasmidprep of HPS AP04 #7-14 & HPS AP10 #7-14 & MDH AP00 #7-14 | ||
+ | |||
+ | {|class="wikitable" | ||
+ | ! construct & clone number !! cryo ID !! plasmid ID !! further procedure | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #7 || #RBVV# || #C4VV# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #8 || #LACA# || #FCOY# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #9 || #E9VF# || #ZNWW# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #10 || #DYDR# || #O3AH# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #11 || #DRNL# || #MVSO# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #12 || #MFPT# || #VOLC# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #13 || #SL4H# || #YNXS# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP04 in pSB1CRDP #14 || #KNX8# || #XOAK# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #7 || #ABAN# || #FMV3# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #8 || #N1RL# || #QPAO# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #9 || #3C6P# || #BRAM# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #10 || #TDYL# || #T4FR# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #11 || #1WXS# || #O4K8# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #12 || #OTE8# || #HRWD# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #13 || #1FX9# || #QZCX# || cancel | ||
+ | |- | ||
+ | | HPS.B1006.AP10 in pSB1CRDP #14 || #86CB# || #YBTE# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #7 || #3QEE# || #TDLS# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #8 || #RFK6# || #8E93# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #9 || #NDM6# || #F69B# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #10 || #PR4Z# || #FN8P# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #11 || #RAYK# || #YF83# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #12 || #HQKY# || #MFCB# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #13 || #R1YY# || #XFLS# || cancel | ||
+ | |- | ||
+ | | MDH.TER.AP00 in pSB1CRDP #14 || #CF4X# || #1131# || cancel | ||
+ | |} | ||
+ | |||
+ | *as problems seem to be mutations in the promoters, building precursors project will be canceled | ||
+ | *we don't have time and money to order new HPLC purified promoters | ||
=References= | =References= |
Latest revision as of 12:53, 18 September 2015