Lab Schedule Overview

Week	Date	Exp-Week	Protocol	Homework due
1	8/25/08	I-1	Outline and Overview Restriction digest Ligation to construct pET28-GFP
2	9/1/08*	I-2	Transformation/plating of ligation Pick colonies (day after lab)	1st Update Expt. I
2	9/1/08*	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	9/8/08	I-3	PCR colony screen Plasmid prepration
3	9/8/08	II-3	A) Observe Synechocystis transformation plate
4	9/15/08	I-4	Restriction digest recombinant plasmids Gel electrophoresis of PCR & digests Transformation of BL21(DE3) Inoculate cultures BL21 (day after lab)
4	9/15/08	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	9/22/08	I-5	Collect and extract cell samples (native)
5	9/22/08	II-5	B) Purification of PCR product
6	9/29/08	I-6	Extract cell samples SDS-PAGE electrophoresis	2nd Update Expt. I
6	9/29/08	II-6	A) Continue segregation for Synechocystis transformants with higher antibiotic B) Restriction digestion of the psbC PCR product and pUC19 plasmid
7	10/6/08	I-7	Metal affinity chromatography of GFP-6His
		II-7	B) Start a plate of wild-type Synechocystis sp. PCC 6803 B) Ligate digests of psbC and pUC19
8	10/13/08	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)	2nd Update Expt. II
9	10/20/08	II-9	B) Prepare recombinant plasmid; submit for sequencing	Report I
9	10/20/08	III-9	Bioinformatics (Part I)
10	10/27/08	II-10	A) Continue segregation for Synechocystis transformants with higher antibiotic
10	10/27/08	III-10	Bioinformatics (Part II)
11	11/3/08	III-11	Genomics (Part I)
11	11/3/08		Lab facilities tour
12	11/10/08**	II-12	A) Small scale DNA preparation from wild- type and the psbC- mutant of Synechocystis B) Gel electrophoresis of recombinant plasmid	3rd Update Expt. II
13	11/17/08	III-13	Genomics (Part II)
13	11/17/08	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	11/24/08	II-14	A) Analysis of PCR products by gel electrophoresis A/B) Check plates for restored photoautotrophic phenotype by transformation of psbC- mutant	Report III
15	12/1/08	Clean-up	Mandatory	Report II (Report III M& T labs)
16	12/8–9/08	Clean-up	For Mon. and Tues. labs only	Report II M& T labs

*The Monday lab will not meet September 1 (Labor Day), and thus will be one week late for all subsequent lab meetings and lab reports.
**The Tuesday labs will not meet November 11 (Veteran’s Day) and thus will be one week late for all subsequent lab meetings and lab reports.

Center for Bioenergy & Photosynthesis

Arizona State University

Box 871604

Room PSD 209

Tempe, AZ 85287-1604

05 September 2008

phone: (480) 965-1963

fax: (480) 965-2747

Contact Webmaster Larry Orr

Accessibility | Privacy