1. FOR Stage 1 Stage 2 Stage 3 J37015 J37016 J37019 J37020 J37025A
Ligation Chart
FOR
Stage 1
Stage 2
Stage 3
J37015
C0261 (Vector) + I13504 (Insert)
PCR Product: 442bp
F2620 (Vector) + (C I13504) (Insert)
Finalised Prey Cell
Riboswitch
Riboswitch (Insert) + (C I13504) (Vector)
PCR Product:
F2620 (Vector) + (RS + C I13504) (Insert)
Issues Talk to john SY
Finalised Prey Cell with Riboswitch
LoxP
Lox Sites (UNKNOWN) + (C I13504) (UNKNOWN)
F2620 (Vector) + (LOX + C I13504) (Insert)
Finalised Prey Cell with Lox Sites
J37016
J37019 (Vector) + I13504 (Insert)
Monocistronic Predator Test Construct
J37019
B0034 (Vector) + C0062 (Insert)
PCR Product: 527bp
R0062 (Vector) + (B C0062) (Insert)
J37020
I13033 (Vector) + I13504 (Insert)
PCR Product: 434bp
J37019 (Vector) + (I I13504) (Insert)
PCR Product: 265bp
Polycistronic Predator Test Construct
J37025A
B0034 (Vector) + J37024 (Insert)
AiiA For Predator
J37024A
J37023 (Insert) + B0015 (Vector)
J37022A
J04500 (Vector) + J37024 (Insert)
AiiA Test Construct
Cre
J04500 (Vector) + Cre (Insert)
(J Cre) (Insert) + B0015 (Vector)
Cre - required for finalised prey cell with LoxP sites
Final Ligation
J37019 (Vector) + J37025 (Insert)
C0261 (24A)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
I13504 (12D)
Cut: X & P
Primer: I13504
F2620 (6B)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
C I13504
Cut: X & P
Primer: I13504
Riboswitch
Cut: Pre-cut
Primer: Riboswitch
C I13504
Cut: E & X
Primer: pSB1A2 Primer (Reverse)
F2620 (6B)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
RS + C I13504
Cut: X & P
Primer: I13504
Depends on PCR which is yet to work in our lab
Lox Sites
Cut: UNKNOWN
Primer: UNKNOWN
C I13504
Cut: UNKNOWN
Primer:UNKNOWN
F2620 (6B)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
LOX + C I13504
Cut: X & P
Primer: I13504
J37019
Cut: S & P
Primer: pSB1(A2/A3/AK3)
I13504
Cut: X & P
Primer: I13504
B0034 (3O)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
C0062 (7A)
Cut: X & P
Primer: C0062 2
R0062 (9G)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
B C0062
Cut: X & P
Primer: C0062 2
I13033 (2H)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
I13504 (12D)
Cut: X & P
Primer: I13504
I I13504
Cut: X & P
Primer: I13504
J37019
Cut: S & P
Primer: pSB1(A2/A3/AK3)
B0034 (3O)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
J37024
Cut: X & P
Primer: UNKNOWN
Waiting on immuno tag
Depends on PCR which is yet to work in our lab
J37023
Cut: S & E
Primer: UNKNOWN
B0015 (1I)
Cut: X & E
Primer: UNKNOWN
J04500 (16P)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
J37024
Cut: X & P
Primer: UNKNOWN
Cre
Cut: X & P
Primer: UNKNOWN
J04500 (16P)
Cut: S & P
Primer: pSB1(A2/A3/AK3)
J37023
Cut: S & E
Primer: UNKNOWN
B0015 (1I)
Cut: X & E
Primer: UNKNOWN
J37019
Cut: S& P
Primer: pSB1(A2/A3/AK3)
J37025
Cut: X &P
Primer: J37024A