Team Hawaii Pcc6803 Electroporation Of Pcc6803 2008 Igem Org
Electroporation of pcc6803 . this experiment can be used to introduce autonomously replicating plasmids to pcc6803. methods . 50 ml linear phase synechocystis at od 730 0.5 collected by centrifugation wash cells three times with 1mm 4 (2 hydroxyethyl) 1 piperazineethanesulfonic acid buffer ph 7.5. Triparental conjugation is a successful method of introducing foreign dna into pcc6803. nine days after plating on bg 11 sp sm, pcc6803 growth was observed at 1 1, 1 10, and 1 100 dilutions of e. coli. no growth was observed for conjugation of pcc6803 with e. coli containing only rp1 (negative control). rp1 itself does not confer sm sp resistance. Grew pcc6803 to od730 = 0.2 and 1.5 (ideal is od730 = 0.5 0.8) transferred 0.75ml cells to sterile eppendorf tube centrifuged 8 min @ 7000g for 8 minutes at room temperature. From 2008.igem.org theory behind tri parental matings "mobilization of a plasmid into a recipient cell is often used for cloning, transposon mutagenesis, or other procedures. Electroporation of pcc6803; completed . pcr amplification of prl1383a; pcc6803 cell density test; pcc6803 cryostock thaw test; effect of co 2 on growth experiment; effect of water motion on clumping of liquid cultures; make competent e. coli; test competent e. coli; initial e. coli transformation; initial biobrick plasmid extraction from filter.
Abstract. we would like to correlate od 750 with specific cell densities (# cells ml) . protocol. plated 200 μl of 1, 10 1, 10 2, 10 5, 10 7, and 10 8 dilutions of pcc6803 on bg 11. Plated synechocystis sp. pcc6803 colonies successfully transformed wwill exhibit a glowing halo of secreted gfp. transformed synechocystis sp. pcc6803 grown in liquid media will result in fluorescent culture media. the efficacy of the signal peptides in transporting gfp into the extracellular media can be measured using a spectrofluorometer. Chemical 2. electroporation 3. conjugation (mr) 2008.06.12 final draft of proposal due 2008.06.23 transform pcc6803 w prl1383a 2008.06.24 first batch of primers ordered 2008.06.24 biobrick parts list completed 2008.06.24 rp1 and rp4 orit are the same 2008.08.13 replaced prl1383a mcs with biobrick mcs (gk).
Team Hawaii Demonstrate 2018 Igem Org