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Project Information


Reference Number 20000133
Project Title Laboratory Exercises in NIMBB Courses: I. MBB 120: Animal Tissue Culture; II. MBB 120: Plant Callus Induction; III. MBB 140: Transformation of DH5 with pUC 18; IV. MBB 240: Advances in Genetic Manipulation; V. MBB 292: Techniques in Molecular Biology II
Project Type Contained Use (Laboratory)
Status Terminated
Name of Institution National Institute of Molecular Biology and Biotechnology (NIMBB), UP Diliman, Quezon City
Cooperating Institution N/A
Supervising IBC UP Diliman-IBC
Project Leader(s) Virginia D. Monje, Professor 6
Experimental Facility/Site NIMBB-UP Diliman, Quezon City
Purpose / Objectives I. 1) To familiarize the students with the techniques in establishing a primary animal cell culture;
2) To measure the viability of the cells involved;
II. 1) To introduce the students to the steps in callus induction in seeds;
2) To determine the effect of environmental factors on callus growth;
III. 1) To train the students on basic techniques in the isolation of plasmids, preparation of vectors and inserts, and transformation.
IV. 1) To exercise the DNA inserts of pAcUW51 and reinsert it in common vector pUC19;
2) To transform E. coli DH5 with the constructed vector;
3) To screen for transformants using Polymerase Chain Reaction (PCR)
V. 1) To perform transformation in prokaryotic host cells using a GFP gene cloned in an expression vector;
2) To express the GFP by arabinose operon induction;
3) To extract and purify the induced GFP using enzyme lysis and chromatography respectively.
Biosafety measures 1. Disposal of cell cultures, used media and other contaminated apparatus will be done by autoclaving at 15 psi (121° C) for 45 mins.
2. Containment level P1
Conditions for Approval N/A
Date Received 2000-04-13
Date of Approval 2000-00-05

 


Regulated Materials


Regulated Materials Plasmids- pUC 18, pUC 19, pGLO, pAcUW 51; Gene Inserts- GFP gene, Ara C, CaMV35S-GUS-NOS cassette;Host Systems- E. coli strains HB101 and DH5
Source N/A
Subject Request of Dr. Elma Llaguno for proposal endorsement
Requesting Party Dr. Elma Llaguno, Chairman, UP Diliman IBC
Action of NCBP Approved for implementation under the AMOR Research Program