Lab Schedule Overview

Week	Date	Exp-Week	Protocol	Homework due
1	8/21/2007	I-1	Outline and Overview Restriction digest Ligation to construct pET28-GFP
2	8/28/2007	I-2	Transformation/plating of ligation Pick colonies (day after lab)	1st Update Expt. I
2	8/28/2007	II-2	Outline and Overview A) Transformation/plating of wild-type Synechocystis with the pKCP43 plasmid; plate cells on agar w/antibiotic B) PCR wild-type Synechocystis psbC gene
3	9/4/2007	I-3	PCR colony screen Plasmid prepration
3	9/4/2007	II-3	A) Observe Synechocystis transformation plate
4	9/11/2007	I-4	Restriction digest recombinant plasmids Gel electrophoresis of PCR & digests Transformation of BL21(DE3)	2nd Update Expt. I
4	9/11/2007	II-4	A) Segregation of Synechocystis transformants: Transfer to a new plate with higher antibiotic concentration B) Gel electrophoresisof PCR products	1st Update Expt. 2
5	9/18/2007	I-5	Inoculate cultures BL21 (day before lab) Subculture and induce BL21 Collect cell samples
5	9/18/2007	II-5	B) Purification of PCR product
6	9/25/2007	I-6	Extract cell samples SDS-PAGE electrophoresis	3rd Update Expt. I
6	9/25/2007	II-6	A) Continue segregation for Synechocystis transformants with higher antibiotic B) Restriction digestion of the psbC PCR product and pUC19 plasmid
7	10/2/2007	I-7	Metal affinity chromatography of GFP-6His	2nd Update Expt. II
7	10/2/2007	II-7	B) Start a plate of wild-type Synechocystis sp. PCC 6803 B) Ligate digests of psbC and pUC19
8	10/9/07	II-8	A) Continue segregation for Synechocystis transformants with higher antibiotic B) Transform E. coli with ligation mix; plate cells B) Start cultures of E. coli transformants (one day prior to week 10 lab period)	Report I
9	10/16/2007	II-9	B) Prepare recombinant plasmid; submit for sequencing	3rd Update Expt. II
9	10/16/2007	III-9	Intoduction to bioinformatics (Part I)
10	10/23/2007	III-10	Intoduction to bioinformatics (Part II)
10	10/23/2007	II-10	A) Continue segregation for Synechocystis transformants with higher antibiotic
11	10/30/2007	III-11	Intoduction to bioinformatics (Part III)	4th Update Expt. II
11	10/30/2007		Lab facilities tour
12	11/6/2007	II-12	A) Small scale DNA preparation from wild- type and the psbC- mutant of Synechocystis B) Gel electrophoresis of recombinant plasmid
13	11/13/2007	III-13	introduction to bioinformatics (Part IV)
13	11/13/07	II-13	A/B) Transform psbC-Synechocystis with recombinant plasmid and wild-type DNA A) PCR of DNA from wild type and the psbC- transformant of Synechocystis
14	11/20/07	III-14	introduction to bioinformatics (Part V)
14	11/20/07	II-14	A) Analysis of PCR products by gel electrophoresis A/B) Check plates for restored photoautotrophic phenotype by transformation of psbC- mutant
15	11/27/2007	III-15	introduction to bioinformatics (Part VI)	Report II
15	11/27/2007	Clean-up	Mandatory
16	12/04/07	turn in	Lab Report III at lecture time	Report III

Center for Bioenergy & Photosynthesis

Arizona State University

Box 871604

Room PSD 209

Tempe, AZ 85287-1604

28 August 2007

phone: (480) 965-1963

fax: (480) 965-2747

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