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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
