Melbourne/Lab GV7

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Vector Insert LIGATION Transform R= ESULT  PICKS Plate
Bba_J61035 digested Antarctic phosphatas= e heat
inactivated		 gel purified Source dnpI
digest		 gel purified digested Alkaline
Phosphatase		 Antarctic phosphatas= e heat
inactivated		 gel purified Type vector qty insert qty amount Target cells #colonies
day 1
BU PCR &X,P (1hr) yes extra digestion in gel (no good)
P48J(1) S,P 80deg20m yes LL PCR 1hr yes X,P 2hr 80deg20m yes 5 min 4uL 4uL all XL-10 gold (45uL) 10 (day2)
P48J(1) S,P 80deg20m yes BU PCR 1hr 5 min 4uL 4uL all XL-10 gold (45uL) 10 (day2) BUA,BUB (21/9/07)
P48J(1) S,P 80deg20m yes LL PCR 1hr yes X,P 2hr 80deg20m yes 5 min 4uL 25uL all XL-10 gold (45uL) 10 (day2)
P48J(1) S,P 80deg20m yes BU PCR 1hr 5 min 4uL 25uL all XL-10 gold (45uL) 10 (day2) BUC,BUD (21/9/07)
P48J(1) S,P 80deg20m yes 4uL all XL-10 gold (45uL) lots of colonies
P48J(1) S,P 1hr 80deg20m yes LLPCR2 1h 30m yes X,P 2hr 80deg20m yes 5 min 2uL 8uL(8ng/ul) 20uL(22ng/ul) Xl-10gold (20uL) 100
P48J(1) S,P 1hr 80deg20m yes BUPCR2 1h 30m yes X,P 2hr 80deg20m yes 5 min 2uL 8uL(8ng/ul) 20uL(22ng/ul) Xl-10gold (20uL) 15
P48J(1) S,P 1hr 80deg20m yes 5min 2uL 20uL Xl-10gold (20uL) covered
P48J(1) S,P 1hr 80deg20m yes LLPCR2 1h 30m yes X,P 2hr 80deg20m yes 16hr 16deg 2uL 20uL Xl-10gold (20uL) 1000 LLA,B,C,D,E
P48J(1) S,P 1hr 80deg20m yes BUPCR2 1h 30m yes X,P 2hr 80deg20m yes 16hr 16deg 2uL 20uL Xl-10gold (20uL) 300 BUA,B,C,D,E 25/9/07 digested in E,X,S
P48J(1) S,P 1hr 80deg20m yes 16hr 16deg 2uL 8uL(8ng/ul) 20uL(22ng/ul) Xl-10gold (20uL) 300
P48J(1) S 1hr complete digest
P48J(1) P 1hr incomplete digest
P48J(1) P 1hr (in B3) complete digest
P48J(1) S,P overnight yes 2uL 45
S,P overnight yes 5 min 2uL 23
S ovnight B2-> P (B3) 2hr yes 2uL 48
S ovnight B2-> P (B3) 2hr yes 5 min 2uL 28
S only yes 5 min 2uL 3000
S only yes 2uL 550
PUC18 ref 5 min 0.5uL 84
PUC18 ref ovnt 0.5uL 61
PUC18 ref 0.5uL 830
P48J(1) S ovnight B2-> P (B3) 2hr yes BUPCR4 1hr yes X,P 2h in (B2) then 2h in (B3) 16hr 16degC 1uL 12ul 20ul Xl-10gold (30uL) fewer 1:12A,B (5/10/07)
P48J(1) S ovnight B2-> P (B3) 2hr yes BUPCR4 1hr yes X,P 2h in (B2) then 2h in (B3) 16hr 16degC 2uL 8ul 20ul Xl-10gold (30uL) fewer 2:8A,B
P48J(1) S ovnight B2-> P (B3) 2hr yes BUPCR4 1hr yes X,P 2h in (B2) then 2h in (B3) 16hr 16degC 2ul 2ul 20ul Xl-10gold (30uL) covered 2:2A,B
P48J(1) S ovnight B2-> P (B3) 2hr yes BUPCR4 1hr yes X,P 2h in (B2) then 2h in (B3) 16hr 16degC 1ul 4ul 20ul Xl-10gold (30uL) covered 1:4A,B
P48J(1) S ovnight B2-> P (B3) 2hr yes BUPCR4 1hr yes X,P 2h in (B2) then 2h in (B3) 16hr 16degC 2ul 20ul Xl-10gold (30uL) covered
P320I X,P 5hrs yes 1hr 5min 1ul 6ul 20ul Xl-10gold (30uL) fewer
P320I X,P 5hrs yes 1hr 5min 2ul 20ul Xl-10gold (30uL) covered
P48J(2) ES (5h40m) BUPCR5 1 hr yes ES 2h 2uL(11ng/ul) 2ul(8ng/ul) 2uL Xl-10gold (45uL) 400 60,61,62 1'
P48J(2)(252ng/u= l) S (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min BUPCR5 1 hr yes X,S 2h 0.4uL(63ng/ul) 3uL(8ng/ul) 2uL Xl-10gold (45uL) 96 63,64 2
P320I(1)(154ng/= ul) ES  (5h40m)<= /td> BUPCR5 1 hr yes ES 2h 2uL(9ng/ul) 3ul(8ng/ul) 2uL Xl-10gold (45uL) 37 3
P48J(2) ES  (5h40m)<= /td> LLPCR3 1 hr yes ES 2h 1uL (11ng/ul) 4ul(5ng/ul) 2uL Xl-10gold (45uL) 120 50,50,52 4
P48J(2) S  (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min LLPCR3 1 hr yes X,S 2h 0.3uL(63ng/ul) 4uL(5ng/ul) 2uL Xl-10gold (45uL) 35 53,54 5
P320I(1) ES  (5h40m)<= /td> LLPCR3 1 hr yes ES 2h 2uL(9ng/ul) 4uL(5ng/ul) 2uL Xl-10gold (45uL) 33 57,58,59 6
P48J(2) ES  (5h40m)<= /td> 2uL (11ng/ul) 2uL Xl-10gold (45uL) 60 71,72,73 7
P48J(2) S  (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min 0.6uL(63ng/ul) 2uL Xl-10gold (45uL) 35 8
P320I(1) ES  (5h40m)<= /td> 3uL(9ng/ul) 2uL Xl-10gold (45uL) 4 9
P48J(2) ES (5h40m) BUPCR5 1 hr yes ES 2h 2uL(11ng/ul) 2ul(8ng/ul) 2uL DH5a(50uL) 11 1'
P48J(2)(252ng/u= l) S (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min BUPCR5 1 hr yes X,S 2h 0.4uL(63ng/ul) 3uL(8ng/ul) 2uL DH5a(50uL) 18 65,66,67,68,69,70 2
P320I(1)(154ng/= ul) ES  (5h40m)<= /td> BUPCR5 1 hr yes ES 2h 2uL(9ng/ul) 3ul(8ng/ul) 2uL DH5a(50uL) 10 3
P48J(2) ES  (5h40m)<= /td> LLPCR3 1 hr yes ES 2h 1uL (11ng/ul) 4ul(5ng/ul) 2uL DH5a(50uL) 0 4
P48J(2) S  (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min LLPCR3 1 hr yes X,S 2h 0.3uL(63ng/ul) 4uL(5ng/ul) 2uL DH5a(50uL) 5 55,56 5
P320I(1) ES  (5h40m)<= /td> LLPCR3 1 hr yes ES 2h 2uL(9ng/ul) 4uL(5ng/ul) 2uL DH5a(50uL) 4 6
P48J(2) ES  (5h40m)<= /td> 2uL (11ng/ul) 2uL DH5a(50uL) 66 7
P48J(2) S  (5h40m) 15m @ 37deg 1hr<= /td> 80deg 20min 0.6uL(63ng/ul) 2uL DH5a(50uL) 0 8
P320I(1) ES  (5h40m)<= /td> 3uL(9ng/ul) 2uL DH5a(50uL) 11 9

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