"
Ryan Quan's notebook
From 2009.igem.org
10/9/09
-miniprepped cultures from 10.7.09 and sent in for sequencing
10/7/09
-set up cultures for 3
10/2/09
-checked sequences
9/30/09
-miniprepped cultures for 1,3,4,5,6,8 -sent in sample for sequencing
9/28/09 -ran gel for PCRs -set up cultures for 1,3,4,5,6,8
9/25/09 -Set up PCR for biobrick parts 2 and 9 with elongase
9/18/09 -ran PCR again for 2 and 9 using KOD -ran gel for PCR
9/11/09 -ran gel again for PCRs
9/9/09 -ran a gel for PCRs -set up BPGW for PCRs -xformed into dh5alpha
9/4/09 -set up PCR for biobricks -DPN1 treated -PCR purified
8/26/09 -set up test digest for ligations of 347-92,93 and FRB-FKBP-FKBP -ran a gel for test digests
8/21/09 -made movies of ho46,320,331,332,333,334,335
8/19/09 -set up cultures for ligations of pho347-93,92 and FRB-FKBP-FKBP
8/13/09 -heat inactivated Aar1 digest -ran gel for digest -gel purified -set up ligations for pho347 w/ 92,93 and AB FRB, BC FKBP, CD FKBP
8/12/09 -miniprepped pho347 -set up Aar1 digest of pho347 o/n
8/11/09 -miniprepped LR cultures -sent in for sequencing (LRs) -made movie of ho46,320,331,332,333,334 -set up culture for pho347
8/10/09 -froze ho335,336 -set up cultures for LRs of pho203-pho275 to pho203-pho382 -xformed pho347 for plasmid amp
8/7/09 -made movies of ho46,180,181,182,227 -checked sequences
8/6/09 -sent in samples for sequencing from 8/5/09 test digest -set up LR rxn for pho375,376,377,378,379,380,381
8/5/09 -set up LRGW for good clones of ligations -froze ho46 w/ pho304 --ho3 -miniprepped cltures from 8/4/09 -set up test digest for minipreps -ran a gel for test digests
8/4/09 -set up for movies of ho46,179,180,181,182,227 -set up cultures for 347-50-60-92-93-143-353 -identified good clones from ligations
8/3/09 -sent in ligations from 7/31/09 for sequencing -set up fruiting bodies for ho46,180,181,182,193,201,203,227
7/31/09 -set up test digest for ligations -ran a gel for tests digests
7/29/09 -miniprepped plasmid amps and BPGW -xformed into dicty ho46: pho303,304,309,311,314,317,322
7/28/09 -set up to take movies for ho46,159,164,171,178,179,193,201,203 -set up cultures for ligations, bpgw and plasmid amp
7/24/09 -set up PCR for new part w/ ho388-ho389 primers on pho337 -set up ligations for PAKAN w/ all Cd's -ran gel for PCR of pho337 w. ho388-389 -set up GWBP for PCR product -xformed ligations and BPGW and plasmid amps-pho 303,304,309,317,322
7/23/09 -recorded movies of ho46,159,164,171,178,179,193,203 -ran Aar1 digests on a gel of 1,6,7,8,60,50,66,97 -gel purified fragments
7/22/09 -set up Aar1 digest for 1,6,7,8,60,66,50,97
7/21/09 -miniprepped cultures for GWBP 1,6,7,8 -ran a gel for Aar1 digest of GWBP parts: 2,3,4,5 -gel purified fragments -xformed pho 223, 228, 233, 341, 342, 343 into ho2 -sent in sequencing of minipreps od 1,6,7,8
7/20/09 -set up cultures of pho223,233,228 and PCRs of new parts 1,6,7,8 -set up aar1 digest of 2,3,4,5 pcrs (fkbp and frbs)
7/17/09 -rna gel of pcrs to check new parts--success -xformed 32 of 71 plasmids into ho2 (Pi3k1/2 minus) -GWBP pcrs of pakan, pipkj, and pten -froze ho201
7/16/09 -miniprepped cultures -sent sequencing for Pd, PD*, pipk-2, pho112 -set up pcr for pakan and pipkj again -set up fruiting body assay for ho171, 159, 46 -set up pcr for pakan, pipkj and new piece, pten (1-16 delta) -ran gel for PCR--success -xformed pho223 and 233 again -set up culture for bp reactions of new parts -froze ho193
7/15/09 -set up pcr for new parts: PakaN, FKBP, FKBP, FKBP, FRB, INP54, PIPKJ -ran pcrs on gel to see if it worked--pakan and pipkj did not work -set up pcrs for gwbp -xformed into bacteria -set up cultures of plasmid amp
7/14/09 -added hygromycin to xformations -checked sequences, alw150 is actually ~250bp, not ~1900. -xformed plasmids for amplifications
7/13/09 -xformed pho303,304,309,311, 317 into ho46 -sent in alw150 from ben for sequencing
7/10/09 -ran gel for colony pcr--not enough template -set up test digest of same samples of colony pcr -ran gel for test digests--success
7/9/09 -ran gel for Aar1 digests--success -gel purified frags -set up ligations for alw150 w/ pho50 -set up colony pcr for alw150 w/ pho 93,92 -set up cultures for colony pcr samples -xformed ligations -set up strains for fruiting bodies HO 178, 171, 159, 172, 46
7/8/09 -miniprepped cultures -set up pvu1 digests -set up Aar1 digests for ALW150 and pho50 -ran gel for test digests -pho92 frag is good, most worked. -set up ligations of alw150 with pho92 and pho 93 -sent in sequences -heat inactivated Aar1 digests -xformed ligations
7/7/09 -set up cultures of ligations -Set up Aar1 digests of pho50,92,93,99
7/6/09 -xformed pho268, 267, 272 and 271 into ho46 -set up ligations of pho55, 66, 99, 132, 143, 144 into pho189 -set up GWLR for pho 255,264,259,256,262,260,263 into pho203 -xformed ligations and gateways into dh5a
7/2/09 -miniprepped cultures for rassl -sent out rassl for sequencing -created bc linkers with oligos -CIP'd Aar1 digest of pho189 -created BC linker again -pcr purified Aar1 digest w/ cip -ran pho189 digests on a gel --success
7/1/09 -set up digest of pho97 and pho189 with nru1 and bamh1/xba1 respectively -pcr purified digests -set up aar1 digest for both -set up cultures of RASSL in pDONR221 from GWBP
6/30/09 -Set up cultures of GWLR reactions in destination vectors and cultures of storage vectors for Aar1
6/29/09 -CAR1 was good but RASSL was not -Set up PCR with new RASSL template -Ran a Gel for PCR -RASSL is good
6/16/09 -Sent in RASSL and CAR1 in PDONR221 for sequencing
6/15/09 -Set up cultures for RASSL and CAR1 in PDONR221
6/11/09 -Ran a gel for PCR's -Gel purified both fragments -Did a gateway BP reaction
6/10/09 -Set up PCR to amplify RASSL and CAR1
