FOR Stage 1 Stage 2 Stage 3 J37015 J37016 J37019 J37020 J37025A

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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) + (C0261 + I13504) (Insert) Finalised Prey Cell Riboswitch Riboswitch (Insert) + (C0261 + I13504) (Vector) PCR Product: F2620 (Vector) + (RS + C0261 + I13504) (Insert) Issues Talk to john SY Finalised Prey Cell with Riboswitch LoxP Lox Sites (UNKNOWN) + (C0261 + I13504) (UNKNOWN) F2620 (Vector) + (LOX + C0261 + 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) + (B0034 + C0062) (Insert) J37020 I13033 (Vector) + I13504 (Insert) PCR Product: 434bp J37019 (Vector) + (I13033 + 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) (J04500 + 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) C0261 + I13504 Cut: X & P Primer: I13504 Riboswitch Cut: Pre-cut Primer: Riboswitch C0261 + I13504 Cut: E & X Primer: pSB1A2 Primer (Reverse) F2620 (6B) Cut: S & P Primer: pSB1(A2/A3/AK3) RS + C0261 + I13504 Cut: X & P Primer: I13504 Depends on PCR which is yet to work in our lab Lox Sites Cut: UNKNOWN Primer: UNKNOWN C0261 + I13504 Cut: UNKNOWN Primer:UNKNOWN F2620 (6B) Cut: S & P Primer: pSB1(A2/A3/AK3) LOX + C0261 + 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) B0034 + C0062 Cut: X & P Primer: C0062 2 I13033 (2H) Cut: S & P Primer: pSB1(A2/A3/AK3) I13504 (12D) Cut: X & P Primer: I13504 I13033 + 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