Team:CGU Taiwan/Notebook

Description

Yeast With IL-8 Receptor

Toehold Switch As RNA Senor

Prototype Design

GPCR

2015.7.1

Operator: Wan yun, Jinting , Jinye
Goal:
  1. Extraction of gDNA of Far1∆::KANMX strain
  2. Detection the concentration of fast extracted gDNA of ∆Far1 strain
  3. PCR gDNA of Far1∆ ::KANMX
  4. Electrophoresis to check PCR product
Experiment steps:
< Extraction of gDNA of Far1∆::KANMX strain>
Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

Name	Pur.	Seq.(5’-3’)	Size (mer.)	MW (g/mol)	Tm (℃)	GC (%)	Nmole	μl for 100μM
dFAR1 F’	Desalt	ggTTTTgTTAggCgggCAAg	20	6244.1	53.8	55	21.25	212.50
dFAR1 R’	Desalt	CATTAACTgCTATTTACgACgC	22	6669.4	51.1	41	17.12	171.20

2. PCR program

Step	Temperature	Time
Step1	95℃	5min
Step2	95℃	30s
Step3	52℃	30s
Step4→step2 for 30 cycle	72℃	2min
Step5	72℃	5min
Step6 (hold on)	10℃	1h

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20

RNA

2015.7.1

Operator: Wan yun, Jinting , Jinye Goal: 1. Extraction of gDNA of Far1∆::KANMX strain 2. Detection the concentration of fast extracted gDNA of ∆Far1 strain 3. PCR gDNA of Far1∆ ::KANMX 4. Electrophoresis to check PCR product Experiment steps: < Extraction of gDNA of Far1∆::KANMX strain> Consult the protocol

Strains	260/280	260/230	C(ng/μl)
Far1∆::KANMX strain	1.62	0.97	44.7

1. Design of primers

					Tm (℃)	GC (%)	Nmole	μl for 100μM
						55	21.25	212.50
						41	17.12	171.20