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00:37, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week20 ‎ (Created page with " ===pRB23-T variants=== We amplified pRB23ΔccdB with KH3/4 in 2x 50 ul Phusion HF reactions and did a gel extraction. <gallery widths="400px" heights="300px"> file:201309 KH3/...")
00:36, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week19 ‎ (Created page with " ===pRB15-18=== The first mini ===Screening delC=== {|class="wikitable" |+Table 15.x Screening pRB18 |- !Template!!Primer!!Comment |- |rowspan="2"|TOP10+pSB3K3||RB64/VR||rowsp...")
00:35, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week18 ‎ (Created page with " ===Preparation for T-Domain exchange=== ====Assembly of pRB19==== PCR indC(ccdB) RB27/28 and pSB1C3 RB21/22 25 ul Phusion Flash HF MM 2x; 5 ul Primer 10 uM each; template ac...")
00:34, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week17 ‎ (Created page with " ===Preparation for T-Domain exchange=== ====Fragment Amplification==== {|class="wikitable" |+Table 12.x PCR for T/TE-Domain exchange with CPEC. Phusion Flash HF |- |colspan="...")
00:34, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week16 ‎ (Created page with " ===Preparation for T-Domain exchanges=== We assemble pRB14, which is a version of pRB3, in which the T-Domain is exchanged by the whole insert of the pDONR vector. Since pRB13 ...")
00:33, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week15 ‎ (Created page with " ===CPEC Conditions Assay (Ralf)=== We performed CPEC assembly of pRB13 with eight different CPEC conditions to see whether we get difeerent results. '''Table 11.1 CPEC mix for...")
00:32, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week14 ‎ (Created page with " ===Konrad=== ====validation of ccdB constructs==== * MP of pKH2(ccdB) constructs from independent colonies: 3rd and 4th were used according to colony PCR result ** (elution in ...")
00:32, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week13 ‎ (Created page with " ===Indigoidine production Konrad (Konrad)=== [[File:20130714-f7-delRest.png|400px|thumb|right|PCR for amplification of pSB1C3: ('''left''') first analytical gel; ('''right'''...")
00:31, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week12 ‎ (Created page with " ===Indigoidine production pKH and pMM (Konrad)=== ==== IPTG Induction ==== [[File:080713-IPTG-induction.png|500px|thumb|right|Induction of 2 days old cultures with/-out 1 mM IP...")
00:31, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week11 ‎ (Created page with " ===Indigoidine production with pKH1 II (Konrad)=== ==== fragment amplification ==== ==> f1: bpsA (all) * 14.6 µl H2O * 25 µl Phusion Flash * 2x 0.5 µl Primer (NI01,NI06)...")
00:31, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week10 ‎ (Created page with " In parallel to the experiments with the pMM-plasmids (Konrad) we start to get the native bpsA from Streptomyces lavendulae lavendulae DSMXXXX (Ralf). ===Indigoidine productio...")
00:30, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week9 ‎ (Created page with " We want to assemble pKH1, which is a pSB1C3-derived plasmid containing pSB1C3-lacPromotor-BBa_B0034-bpsA(pMM64)- BBa_B0034-svp(pMM65). Single fragments are amplified and clone...")
00:30, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week8 ‎ (Created page with " ===Indigoidine production with pMM-plasmids IV (Ilia)=== * inoculate 5 ml LB + Kan with TOP10-pMM65 * miniPrep of pMM65 -> 23 ng / µl * [[Preparing_glycerol_stocks|prepare TOP...")
00:29, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week7 ‎ (Created page with " We received the E. coli BAP1-strain which is commonly used for heterologuos expression of NRPS. It carries sfp, a 4'-Phosphopanthetheinyl-Transferase from Bacillus subtilis t...")
00:28, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week6 ‎ (Created page with " Transformed Rosetta with pMM64 and pMM65 didn't show a blue phenotype. Transformation will be repeated. ===Indigoidine production with pMM-plasmids II (Ilia)=== * one blue colon...") (top)
00:28, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week5 ‎ (Created page with " We got two plasmids from the Fussenegger group at the ETH Zurich containing a codon optimized version of bpsA (pMM64) and svp (pMM65), respectively, for expression in HEK 297-...") (top)
00:26, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week23 overview ‎ (Created page with " ===Domain Shuffling – Indigoidine Synthetase Conversions=== Besides the delH4 T-domain only the plu2642 T-domain resulted in a functional indigoidine synthetase when intro...")
00:25, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week22 overview ‎ (Created page with " ===T-domain shuffling=== Last week's co-transformation experiments showed that exchanging the indC-T-domain with the bpsA T-domain results in a functional indigoidine synthe...")
00:25, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week21 overview ‎ (Created page with " ===BioBricks for the Registry=== This week we prepared our parts for submission to the registry. We submitted the coding sequences of indC, sfp, svp, entD and delC, as well ...")
00:25, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week20 overview ‎ (Created page with " ===T-Domain Shuffling=== We prepared pRB23-T1 to -TTE19 and performed transformations in ''E. coli'' TOP10 as well as co-transformations with the four PPTase...")
00:24, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week19 overview ‎ (Created page with " ===Domain Shuffling – PPTase Plasmids=== The PPTase plasmids pRB15-18 have been reassembled with pSB3K3 from a different plate of the partsdistribution,...")
00:24, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week18 overview ‎ (Created page with " ===T-Domain Shuffling=== We again assembled pRB19 from pRB14 and did all T-domain-exchanges in this plasmid. After we picked and preppe...")
00:23, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week17 overview ‎ (Created page with " ===T-Domain Shuffling=== We started inserting new T-Domains into indC to check their functionality. In total we first assembled 19 variants of pRB19, whic...")
00:22, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week16 overview ‎ (Created page with " ===T-Domain Shuffling – PPTase Plasmids=== This week we wanted to prepare plasmids for the T-Domain exchange experiments. We assembled four plasmids [[Plasmids#pRB15|pRB15...")
00:22, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week15 overview ‎ (Created page with " === Indigoidine Production – Testing PPTases === pRB3 contains indC and sfp and results in a blue phenotype after tranformatio...")
00:22, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week14 overview ‎ (Created page with " ===T-Domain Shuffling – Assembly of ccdB Plasmids=== In the subsequent weeks we wanted to exchange the indC T-domain with several T-domains of other NRPS modules. These ar...")
00:21, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week13 overview ‎ (Created page with " ===Indigoidine Production – Testing PPTases=== We wanted to test various PPTases on their functionality in activating the indigoidine synthetases indC and bpsA. The plasmi...")
00:20, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week12 overview ‎ (Created page with " ===Indigoidine Production - indC=== Since ''S. lavendulae'' DSM40708 does not carry the bpsA gene, we used indC from ''Photorhabdus luminescens laumondii'' TT01 instead (<b...")
00:20, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week11 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== ''Streptomyces verticillus'' ATCC15003 was cultivated and used for amplification of the svp PPTase. We tried to extract bpsA from ''S. la...")
00:20, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week10 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== For the PCR amplification of native bpsA, ''Streptomyces lavendulae subsp. lavendulae'' DSM40708 was grown on GYM agar plates. We wanted ...")
00:19, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week9 overview ‎ (Created page with " ===Indigioidine Production – bpsA=== This week we assembled our first [http://parts.igem.org/Part:pSB1C3 pSB1C3] derived plasmid pKH1, which carries both...")
00:19, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week8 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== We performed PCR screenings of the pMM64 (bpsA) and pMM65 (svp) plasmid for validation. The PCR screenings showed that the pMM-...")
00:18, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week7 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== Since we couldn't observe indigoidine production using pMM64 and pMM65 in the previous two weeks we...")
00:18, 5 October 2013 (diff | hist) Nm Team:Heidelberg/Templates/Indigoidine week6 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== This week we repeated the co-transformation of both the ''E. coli'' Rosetta and the BAP1 strain with the bpsA ([[Plasm...")
00:17, 5 October 2013 (diff | hist) N Team:Heidelberg/Templates/Indigoidine week5 overview ‎ (Created page with " ===Indigoidine Production - bpsA=== At the beginning of our wetlab phase, we wanted to transform ''E. coli'' cells with plasmids containing an [[IndPD#ind_synthetase|indigoi...")
00:03, 5 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
00:02, 5 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
00:00, 5 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:53, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:41, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:34, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:25, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Del week15 overview ‎ (top)
23:21, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Methods ‎
23:20, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Methods ‎
23:17, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:14, 4 October 2013 (diff | hist) m Team:Heidelberg/Templates/Materials list ‎
23:13, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:13, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:12, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎
23:10, 4 October 2013 (diff | hist) Team:Heidelberg/Templates/Materials list ‎

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