Team:NCTU Formosa/Notebook/test6

9/4

Cultivation of

P_cons+RBS+AseI PCR product-1,2,3,4,7,8 Ar

XhoI+PSB1K3 PCR product-1,2 Kr

PSB1A3 Ar

Cultivation plate test

Ar plate and Kr plate

Digestion of

[PBAN-HindIII] Kr*2 (EP)

[PBAN-NheI]Kr*2 (EP)

[PBAN-KpnI-HF]Kr*2 (EP)

[23101+B0034+PBAN (HindIII)]Ar(ES)

[23101+B0034+PBAN (NheI)]Ar(ES)

PCR of

[J23101+B0034+KpnI-HF] Ar*3

Ligation of

[J23101+B0034+KpnI-HF+PSB1C3]

Electrophoresis of

[PBAN-HindIII] Kr*2 (EP)

[PBAN-NheI]Kr*2 (EP)

[PBAN-KpnI-HF]Kr*2 (EP)

[23101+B0034+PBAN (HindIII)]Ar(ES)

[23101+B0034+PBAN (NheI)]Ar(ES)

[23101+B0034+PBAN (HindIII)]Ar(EP)*2

[23101+B0034+PBAN (NheI)]Ar(EP)

[23101+B0034+PBAN (KpnI-HF)]Ar(EP)*2

[23101+B0034+PBAN (KpnI-HF)]Ar PCR*3

9/5

Electrophoresis of

Well 1 P_cons+RBS+AseI--1 (EP) Ar

Well 2 P_cons+RBS+AseI--2 (EP) Ar

Well 3 P_cons+RBS+AseI--3 (EP) Ar

Well 4 P_cons+RBS+AseI--4 (EP) Ar

Well 5 P_cons+RBS+AseI--7 (EP) Ar （DNA sequencing）

Well 6 P_cons+RBS+AseI--8 (EP) Ar

Well 7 P_cons+RBS+NcoI--2 (EP) Cr （DNA sequencing）

Well 8 P_cons+RBS+Ndel--2 (EP) Cr （DNA sequencing）

Well 9 1 Kb marker

Well 10 P_cons+RBS+Ndel--13 (EP) Cr

Well 11 SacI--3 (XP) Kr

Well 12 SacI--4.1 (XP) Kr

Well 13 SacI--4.2 (XP) Kr

Well 14 XhoI--7 (EP) Kr （DNA sequencing）

Well 15 XhoI--8 (EP) Kr（DNA sequencing）

Well 16 SalI--3 (XP) Kr

Mini-prep of

P_cons+RBS+KpnI-HF-7

P_cons+RBS+KpnI-HF-8

P_cons+RBS+KpnI-HF-9

Digestion of

P_cons+RBS+KpnI-HF-7 (EP)

P_cons+RBS+KpnI-HF-8 (EP)

P_cons+RBS+KpnI-HF-9 (EP)

Cultivation of

P_cons+RBS+KpnI-HF Ar PCR product-7,8,9 Ar

Ligation of

[J23101+B0034+HindIII+B0034+PSB1K3] Kr

[J23101+B0034+HindIII+J61048+PSB1K3] Kr

[J23101+B0034+NheI+B0034+PSB1K3] Kr

[J23101+B0034+NheI+J61048+PSB1K3] Kr

Electrophoresis of digested product of

[P_cons+RBS+KpnI-HF]Ar (EP)*3

9/9

Cultivation of

P_cons+RBS+SacI PCR product-16 Ar

P_cons+RBS+SaII PCR product-17 Ar

RBS+BFP+Ter PCR product-16,17,29,30 Ar

Digestion of

XhoI+PSB1K3 (EP)

9/10

Mini-prep of

RBS+BFP+Ter-16 Ar

RBS+BFP+Ter-17 Ar

RBS+BFP+Ter-29 Ar

RBS+BFP+Ter-30 Ar

P_cons+RBS+Sac-16 Ar

P_cons+RBS+Sal-17 Ar

Digestion of

RBS+BFP+Ter-16 Ar(EP)

RBS+BFP+Ter-17 Ar(EP)

RBS+BFP+Ter-29 Ar(EP)

RBS+BFP+Ter-30 Ar(EP)

P_cons+RBS+SacI-16 Ar(EP)

P_cons+RBS+SalI-17 Ar(EP)

Electrophoresis of

Well 1 Marker

Well 2 P_cons+RBS+SacI-16 dig(EP)

Well 3 P_cons+RBS+SalI-17 dig(EP)

Well 4 RBS+BFP+Ter-16 dig(EP)

Well 5 RBS+BFP+Ter-17 dig(EP)

Well 6 RBS+BFP+Ter-29 dig(EP)

Well 7 RBS+BFP+Ter-30 dig(EP)

Well 8 XhoI-7 dig(XP) Kr

9/11

Digestion of

P_cons+RBS+PBAN(ES)(8 kind except of XhoI)

RBS+BFP+Ter(XP)

Ligation of

P_cons+RBS+XhoI-7 Ar

Cultivation of

P_cons+RBS+AseI-7 Ar

P_cons+RBS+SacI-16 Ar

P_cons+RBS+SalI-17 Ar

P_cons+RBS+NcoI-2 Ar

P_cons+RBS+NdeI-2 Ar

RBS+BFP+Ter-2 Ar

RBS+BFP+Ter-2 Ar

SalI-3 Kr

Ligation of

P_cons+RBS+XhoI-7 Ar

DNA sequencing of

P_cons+RBS+SaeI Ar 8ul VF2

P_cons+RBS+SalI Ar 8ul VF2

RBS+BFP+Ter-29 Ar 8ul VF2 + 8ul VR

RBS+BFP+Ter-30 Ar 5ul VF2 + 5ul VR

Digestion of

P_cons+RBS+HindIII dig(EP)

P_cons+RBS+NheI dig(EP)

P_cons+RBS+NcoI dig(EP)-->9/11 mini

P_cons+RBS+NdeI dig(EP)

P_cons+RBS+SacI dig(EP)

P_cons+RBS+SalI dig(EP)

P_cons+RBS+AseI dig(EP)

P_cons+RBS+KpnI dig(EP)

Ligation of

P_cons+RBS+BFP+Ter-30 Kr

9/12

Transformation of

P_cons+RBS+NheI+RBS+BFP+Ter Kr

P_cons+RBS+NdeI+RBS+BFP+Ter Kr

P_cons+RBS+HindIII+RBS+BFP+Ter Kr

P_cons+RBS+KpnI-HF+RBS+BFP+Ter Kr

P_cons+RBS+AseI+RBS+BFP+Ter Kr

P_cons+RBS+SacI+RBS+BFP+Ter Kr

P_cons+RBS+NcoI+RBS+BFP+Ter Kr

P_cons+RBS+SalI+RBS+BFP+Ter Kr

P_cons+RBS+BFP+Ter Kr

P_cons+RBS+XhoI Ar

Electrophoresis of

Well 1 Marker

Well 2 P_cons+RBS+NdeI dig(EP) Cr

Well 3 P_cons+RBS+NcoI dig(EP) Cr

Well 4 P_cons+RBS+SacI dig(EP) Ar

Well 5 P_cons+RBS+SalI dig(EP) Ar

Well 6 P_cons+RBS+AseI dig(EP) Ar

Well 7 P_cons+RBS+HindIII dig(EP) Ar

Well 8 P_cons+RBS+NheI dig(EP) Ar

Well 9 P_cons+RBS+KpnI dig(EP) Ar

9/13

Ligation of

P_cons+RBS+AseI Kr

P_cons+RBS+NdeI Kr

P_cons+RBS+SalI Kr

P_cons+RBS+SacI Kr

PCR of

P_cons+RBS+HindIII+RBS+BFP+Ter Kr x8

P_cons+RBS+NheI+RBS+BFP+Ter Kr x8

P_cons+RBS+NdeI+RBS+BFP+Ter Kr x8

P_cons+RBS+NcoI+RBS+BFP+Ter Kr x8

P_cons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8

P_cons+RBS+AseI+RBS+BFP+Ter Kr x8

P_cons+RBS+Xhol Kr x8

P_cons+RBS+SalI+RBS+BFP+Ter Kr x16

P_cons+RBS+SacI+RBS+BFP+Ter Kr x16

Electrophoresis of

Gel 1

P_cons+RBS+SalI+RBS+BFP+Ter Kr x16

Gel 2

P_cons+RBS+SacI+RBS+BFP+Ter Kr x16

Gel 3

P_cons+RBS+HindIII+RBS+BFP+Ter Kr x8

P_cons+RBS+NheI+RBS+BFP+Ter Kr x8

Gel 4

P_cons+RBS+NdeI+RBS+BFP+Ter Kr x8

P_cons+RBS+NcoI+RBS+BFP+Ter Kr x8

Gel 5

P_cons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8

P_cons+RBS+AseI+RBS+BFP+Ter Kr x8

Gel 6

P_cons+RBS+Xhol Kr x8

psb1K3 dig(EP)

9/16

Transformation of

psb3K3 *2

psb3C3

Digestion of

psb3K3 dig(EP)

psb1C3 dig(EP)

Cultivation of

P_cons+RBS+PBAN+RBS+BFP+Ter x8

9/17

Mini-prep of

PBAN：XhoI

SalI

SacI

KpnI

Ndel

AseI

NcoI

NheI

HindIII (fail)

P_cons+RBS+PBAN+RBS+BFP+Ter：AseI-4

Ndel-8

HindIII-4 (fail)

SacI-9(fail)

KpnI-1

NcoI-2

NheI-1

Digestion of

PBAN：XhoI

SalI

SacI

KpnI

Ndel

AseI

NcoI

AseI

P_cons+RBS+PBAN+RBS+BFP+Ter：AseI-4

Ndel-8

KpnI-1

NcoI-2

NheI-1

Electrophoresis of

P_cons+RBS+PBAN+RBS+BFP+Ter (Ndel)

P_cons+RBS+PBAN+RBS+BFP+Ter (NcoI)

P_cons+RBS+PBAN+RBS+BFP+Ter (SacI)

P_cons+RBS+PBAN+RBS+BFP+Ter (HindIII)

P_cons+RBS+PBAN+RBS+BFP+Ter (SalI)

P_cons+RBS+PBAN+RBS+BFP+Ter (NheI)

P_cons+RBS+PBAN+RBS+BFP+Ter (KpnI)

P_cons+RBS+PBAN+RBS+BFP+Ter (AseI)

PBAN AseI

PBAN Ndel

PBAN KpnI

PBAN SacI

PBAN SalI

PBAN XhoI

P_cons+RBS+XhoI-1 dig(EP)

P_cons+RBS+Xhol-1 dig(ES)

psb1C3 dig(ES)

Cultivation of

P_cons+RBS+PBAN+RBS+BFP+Ter (SacI)-4.12

P_cons+RBS+PBAN+RBS+BFP+Ter (HindIII)-6

P_cons+RBS+PBAN+RBS+BFP+Ter (SalI)-9

Digestion of

B0034 Kr dig(ES)

P_cons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

P_cons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

NcoI Kr dig(EP)

Ndel Kr dig(EP)

SalI Kr dig(EP)

9/18

Electrophoresis of

Well 1 Marker

Well 2 B0034 Kr dig(ES)

Well 3 P_cons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

Well 4 P_cons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

Well 5 NcoI Kr dig(EP)

Well 6 Ndel Kr dig(EP)

Well 7 SalI Kr dig(EP)

Electrophoresis of

Well 1 Marker

Well 2 B0034 Kr dig(ES)

Well 3 P_cons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

Well 4 P_cons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

Well 5 NcoI Kr dig(EP)

Well 6 Ndel Kr dig(EP)

Well 7 SalI Kr dig(EP)

Mini-prep of

P_cons+RBS+HindIII+RBS+BFP+Ter-6

P_cons+RBS+SalI+RBS+BFP+Ter-9

P_cons+RBS+SacI+RBS+BFP+Ter-4

P_cons+RBS+SacI+RBS+BFP+Ter-11

Ligation of

P_cons+RBS+AseI+RBS+BFP+Ter Cr

P_cons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

NcoI Cr

9/20

Transformation of

AseI mini Cr

Electrophoresis of

Well 1 Marker

Well 2 HindIII-2 Kr dig(EP)

Well 3 SacI Kr dig(EP)

Well 4 P_cons+RBS+HindIII-6+RBS+BFP+Ter Kr dig(EP)

Well 5 P_cons+RBS+SacI-11+RBS+BFP+Ter Kr dig(EP)

Well 6 P_cons+RBS+SacI-9+RBS+BFP+Ter Kr dig(EP)

Well 7 RBS+BFP+Ter Ar dig(XP)

Well 8 P_cons+RBS+NheI+RBS+BFP+Ter Kr dig(EP)

9/21

Electrophoresis of PCR products of

Gel 1

Well 1 Marker

Well 2 3H1

Well 3 3H2

Well 4 3H3

Well 5 3H4

Well 6 3H5

Well 7 3H6

Well 8 Marker

Well 9 6K1

Well 10 6K2

Well 11 6K3

Well 12 6K4

Well 13 6K5

Well 14 6K6

Well 15 3Sl1

Well 16 3Sl2

Well 17 3Sl3

Gel 2

Well 1 Marker

Well 2 6Sl4

Well 3 6Sl5

Well 4 3Nh1

Well 5 3Nh2

Well 6 3Nh3

Well 7 3Nh4

Well 8 Marker

Well 9 3Nh5

Well 10 3Nh6

Well 11 6X1

Well 12 6X2

Well 13 6X3

Well 14 6X4

Well 15 6X5

Well 16 6X6

Well 17 1K1

Gel 3

Well 1 Marker

Well 2 1K2

Well 3 1K3

Well 4 1K4

Well 5 1K5

Well 6 1K6

Well 7 1X1

Well 8 Marker

Well 9 1X2

Well 10 1X3

Well 11 1X4

Well 12 1X5

Well 13 1X6

Well 14 3K1

Well 15 3K2

Well 16 3K3

Well 17 3K4

Gel 4

Well 1 Marker

Well 2 3K5

Well 3 3K6

Well 4 3Sc1

Well 5 3Sc2

Well 6 3Sc3

Well 7 3Sc4

Well 8 Marker

Well 9 3Sc5

Well 10 3Sc6

Well 11 1Sc1

Well 12 1Sc2

Well 13 1Sc3

Well 14 1Sc4

Well 15 1Sc5

Well 16 1Sc6

Well 17 1Nc1

Gel 5

Well 1 Marker

Well 2 1Nc2

Well 3 1Nc3

Well 4 1Nc4

Well 5 1Nc5

Well 6 1Nc6

Well 7 1A1

Well 8 Marker

Well 9 1A2

Well 10 1A3

Well 11 1A4

Well 12 1A5

Well 13 1A6

Well 14 1Nd1

Well 15 1Nd2

Well 16 1Nd3

Well 17 1Nd4

Gel 6

Well 1 Marker

Well 2 1Nd5

Well 3 1Nd6

Well 4 3X1

Well 5 3X2

Well 6 3X3

Well 7 3X4

Well 8 Marker

Well 9 3X5

Well 10 3X6

Well 11 3A1

Well 12 3A2

Well 13 3A3

Well 14 3A4

Well 15 3A5

Well 16 3A6

Cultivation of

KpnI-1

NdeI-1

XhoI-1

SacI-1

AseI-1

P_cons+RBS+HindIII-1

P_cons+RBS+KpnI-4

P_cons+RBS+NheI-4

P_cons+RBS+SalI-5

P_cons+RBS+AseI-5

P_cons+RBS+XhoI-1

P_cons+RBS+SacI-1

P_cons+RBS+KpnI-1+RBS+BFP+Ter

P_cons+RBS+XhoI-6+RBS+BFP+Ter

Digestion of

SalI Kr dig(EP)

Ligation of

NdeI dig(EP)+C backbone

9/22

Transformation of

NheI Cr

P_cons+RBS+NheI+RBS+BFP+Ter Cr

P_cons+RBS+NcoI+RBS+BFP+Ter Cr

P_cons+RBS+Ndel+RBS+BFP+Ter Cr

P_cons+RBS+AseI+RBS+BFP+Ter Cr

Digestion of

HindIII(EP)

Cultivation of

SacI-3 Kr

NcoI-7 Cr

NcoI-8 Cr

P_cons+RBS+SacI-2

P_cons+RBS+XhoI-5+RBS+BFP+Ter

pbs3C5

Ligation of

P_cons+RBS+NcoI+RBS+BFP+Ter Cr

P_cons+RBS+HindIII+RBS+BFP+Ter Cr

P_cons+RBS+SacI+RBS+BFP+Ter Cr

P_cons+RBS+SalI+RBS+BFP+Ter Cr

9/23

Electrophoresis of

Well 1 Marker

Well 2 KpnI-1 Cr dig(EP)

Well 3 NdeI-1 Cr dig(EP)

Well 4 XhoI-1 Cr dig(EP)

Well 5 SacI-1 Cr dig(EP)

Well 6 AseI-1 Cr dig(EP)

Well 7 P_cons+RBS+HindIII-1 dig(EP)

Well 8 P_cons+RBS +KpnI-4 dig(EP)

Well 9 P_cons+RBS+NheI-4 dig(EP)

Well 10 P_cons+RBS+AseI-5 dig(EP)

Well 11 P_cons+RBS+XhoI-1 dig(EP)

Well 12 P_cons+RBS+SacI-1 dig(EP)

Well 13 P_cons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)

Well 14 SalI Kr dig(EP) 9/21

Well 15 SalI Kr dig(EP) 9/19

DNA sequencing of

KpnI-1 Cr dig(EP)

NdeI-1 Cr dig(EP)

XhoI-1 Cr dig(EP)

SacI-1 Cr dig(EP)

AseI-1 Cr dig(EP)

P_cons+RBS+HindIII-1 dig(EP)

P_cons+RBS+KpnI-4 dig(EP)

P_cons+RBS+NheI-4 dig(EP)(fail)

P_cons+RBS+AseI-5 dig(EP)

P_cons+RBS+XhoI-1 dig(EP)

P_cons+RBS+SacI-1 dig(EP)

P_cons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)

Transformation of

P_cons+RBS+HindIII+RBS+BFP+Ter

P_cons+RBS+NcoI+RBS+BFP+Ter

P_cons+RBS+SalI+RBS+BFP+Ter

P_cons+RBS+SacI+RBS+BFP+Ter

9/24

Cultivation of

P_cons+RBS+NheI+RBS+BFP+Ter Cr

P_cons+RBS+AseI+RBS+BFP+Ter Cr

P_cons+RBS+NcoI+RBS+BFP+Ter Cr

P_cons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

psb3C5 Cr

Cultivation of

P_cons+RBS+NheI+RBS+BFP+Ter Cr

P_cons+RBS+AseI+RBS+BFP+Ter Cr

P_cons+RBS+NcoI+RBS+BFP+Ter Cr

P_cons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

psb3C5 Cr

Electrophoresis of

Well 1 Marker

Well 2 SalI Kr dig(EP)

Well 3 P_cons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)

Well 4 P_cons+RBS+SacI Cr dig(EP)

Well 5 NcoI Cr dig(EP)

Digestion of

P_cons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)

P_cons+RBS+SacI-2 Cr dig(EP)

NcoI-8 Cr dig(EP)

SalI-3 Kr dig(EP)

Ligation of

HindIII+psb1C3

PCR of

SacI x8

SalI x5

HindIII x2

NheI x8

Mini-prep and digestion of

P_cons+RBS+AseI+RBS+BFP+Ter

P_cons+RBS+NheI+RBS+BFP+Ter

P_cons+RBS+NdeI+RBS+BFP+Ter

P_cons+RBS+NcoI+RBS+BFP+Ter

psb3C5

NheI

9/25

Electrophoresis and DNA sequencing of

Well 1 Marker

Well 2 NheI (fail)

Well 3 P_cons+RBS+AseI+RBS+BFP+Ter

Well 4 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 5 P_cons+RBS+NdeI+RBS+BFP+Ter (fail)

Transformation of

HindIII

SalI

P_cons+RBS+SacI+RBS+BFP+Ter

P_cons+RBS+SalI+RBS+BFP+Ter

P_cons+RBS+HindIII+RBS+BFP+Ter

PCR of

NcoI x5(9.10.11.12.13)

P_cons+RBS+SacI x5(7.8.9.10.11)

Cultivation of

NheI

P_cons+RBS+SacI-2(9/22)

P_cons+RBS+NcoI-8(9/22)

P_cons+RBS+NheI-4

P_cons+RBS+NdeI+RBS+BFP+Ter x2

P_cons+RBS+NheI+RBS+BFP+Ter x2

9/26

Mini-prep and digestion(EP) of

NcoI

NheI

P_cons+RBS+SacI-2

P_cons+RBS+NheI-4

P_cons+RBS+NdeI-1+RBS+BFP+Ter

P_cons+RBS+NdeI-2+RBS+BFP+Ter

Electrophoresis of

Well 1 Marker

Well 2 P_cons+RBS+NdeI-1+RBS+BFP+Ter

Well 3 P_cons+RBS+NdeI-2+RBS+BFP+Ter

Well 4 NcoI

Well 5 NheI

Well 6 P_cons+RBS+NheI-4

Well 7 P_cons+RBS+SacI-2

Cultivation of

psb3C5 x2

HindIII-3

HindIII-4

SalI-3

SalI-4

P_cons+RBS+HindIII-3+RBS+BFP+Ter

P_cons+RBS+HindIII-4+RBS+BFP+Ter

P_cons+RBS+SalI-3+RBS+BFP+Ter

P_cons+RBS+SalI-4+RBS+BFP+Ter

P_cons+RBS+SacI-3+RBS+BFP+Ter

P_cons+RBS+SacI-4+RBS+BFP+Ter

P_cons+RBS+NcoI-3+RBS+BFP+Ter

P_cons+RBS+NcoI-4+RBS+BFP+Ter

9/28

Electrophoresis of

Well 1 HindIII-3

Well 2 HindIII-4

Well 3 SalI-3

Well 4 SalI-4

Well 5 P_cons+RBS+SacI-3+RBS+BFP+Ter

Well 6 Marker

Well 7 P_cons+RBS+NcoI-3+RBS+BFP+Ter

Well 8 P_cons+RBS+NcoI-4+RBS+BFP+Ter

Well 9 P_cons+RBS+SalI-3+RBS+BFP+Ter

Well 10 P_cons+RBS+SalI-4+RBS+BFP+Ter

Well 11 psb3C5

9/29

Electrophoresis of

Well 1 Marker

Well 2 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 3 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 4 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 5 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 6 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 7 P_cons+RBS+HindIII+RBS+BFP+Ter

Well 8 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 9 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 10 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 11 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 12 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 13 P_cons+RBS+NcoI+RBS+BFP+Ter

Well 14 P_cons+RBS+SalI+RBS+BFP+Ter

Well 15 P_cons+RBS+SalI+RBS+BFP+Ter

Well 16 P_cons+RBS+SalI+RBS+BFP+Ter

Well 17 P_cons+RBS+SalI+RBS+BFP+Ter

Well 18 P_cons+RBS+SalI+RBS+BFP+Ter

Well 19 P_cons+RBS+SalI+RBS+BFP+Ter

Well 20 psb3C5 dig

Well 21 Marker

Ligation of

P_cons+RBS+KpnI + pbs3C5

P_cons+RBS+SacI + pbs3C5

P_cons+RBS+AseI + pbs3C5

P_cons+RBS+HindIII + pbs3C5

P_cons+RBS+NheI + pbs3C5

P_cons+RBS+SalI + pbs3C5

P_cons+RBS+NcoI + pbs3C5

P_cons+RBS+NdeI + pbs3C5

P_cons+RBS+XhoI+RBS+BFP+Ter + pbs1C3

P_cons+RBS+NdeI+RBS+BFP+Ter + pbs1C3

P_cons+RBS+SacI + pbs1C3

P_cons+RBS+NcoI + pbs1C3

DNA sequencing of

SalI-4

HindIII-4

P_cons+RBS+SalI-3+RBS+BFP+Ter

P_cons+RBS+SalI-2

P_cons+RBS+NheI-4

NheI(9/26)

P_cons+RBS+HindIII+RBS+BFP+Ter(9/30)

P_cons+RBS+SalI+RBS+BFP+Ter(9/30)

9/30

PCR and Electrophoresis of

P_cons+RBS+NdeI+RBS+BFP+Ter

NcoI

P_cons+RBS+NheI+RBS+BFP+Ter

P_cons+RBS+XhoI+RBS+BFP+Ter

P_cons+RBS+SacI+RBS+BFP+Ter

Cultivation of

P_cons+RBS+NheI-11+RBS+BFP+Ter

P_cons+RBS+NheI-13+RBS+BFP+Ter

P_cons+RBS+NdeI-3+RBS+BFP+Ter

P_cons+RBS+NdeI-10+RBS+BFP+Ter

P_cons+RBS+XhoI-3+RBS+BFP+Ter

P_cons+RBS+XhoI-5+RBS+BFP+Ter

P_cons+RBS+SacI-2+RBS+BFP+Ter

P_cons+RBS+SacI-11+RBS+BFP+Ter

NcoI-3

NcoI-12