Human Genetic Engineering
Provides information and different resources on Human Genetic Engineering
What is Genetic Engineering
is the science of manipulating an organism's genetic material to modify the proteins it produces. It is used in taking genes and segments of DNA from one species to put them into another.
Genetic Engineering provides a set of techniques to cut DNA either randomly or at a number of specific sites. Once the DNA is isolated one can study its different segments, multiply them up and splice them next to any other DNA of another cell or organism. GE makes it possible to break through the species barrier and to shuffle information between completely unrelated species; for example, to splice the anti-freeze gene from flounder into tomatoes or strawberries, an insect-killing toxin gene from bacteria into maize, cotton or rapeseed, or genes from humans into pig.
Laboratory topics in Genetic Engineering
Introduction to the laboratory, basic equipments and techniques.
Preparation of media and reagents commonly used in molecular biology.
Isolation of chromosomal DNA from natural sources: E. coli, Vibrio fisceri.
Spectrophotometric quantification and analysis of DNA.
Restriction digestion and agarose gel electrophoresis of plasmid DNA and bacteriophage DNA.
Construction of a genomic library from Vibrio fischeri: Ligation of genomic DNA fragments and vector analysis by gel electrophoresis.
Isolation of bioluminescent bacteria from natural sources: (Vibrio fischeri, and shrimps).
Preparation of competent E. coli cells.
Transformation of competent cells.
Plasmid mini-preps from bioluminescent clones.
Restriction digestion of plasmid-mini preps.Analysis of digests by agarose gel electrophoresis.
Southern Blot analysis I: Genomic DNA isolation from new bioluminescent isolates.
Southern Blot analysis II: Restriction digestion of genomic DNA from new isolates. Analysis of digests by agarose gel electrophoresis. Transfer of restriction fragments to membrane.
DNA sequencing; autoradiography; analysis of DNA sequence.
Polymerase Chain Reaction (PCR). - Refer to http://www.unixl.com/dir/molecular_sciences/biochemistry/biochemical_techniques/pcr/ for detail on PCR.
Below is the introduction to some of the instruments used in genetic research.
Instruments used for isolation of chromosomal DNA:
25 Falcon tubes (14 ml).
Ampicillin solution (50 mg/ml in sterile water).
Instruments used for the Spectrophotometric quantification and analysis of DNA:
Zeiss LSM 510 - Obtaining Images- A laser scanning confocal microscopy system.
Zeiss Axiovert 100M.
Fluorescence inverted microscope thats used to obtain series of images.
Used for 3D Reconstruction of images:
3.UNIX/IRIX Bitplane Imaris 2.7
4.Bitplane Imaris 3.0
Hardwares used for 3D Reconstruction of images:
1.Graphic Workstation PC/Intel.
Instruments used for Restriction digestion plasmid DNA:
Incubator, containers, pipette.
Instruments used for Ligation of genomic DNA fragments.
Instruments used for DNA Sequencing:
4 ABI 373
2 ABI 3100
1 ABI 3700
1 APB Megabase 1000
Instruments used in Peptide Synthesis:
1 Advanced Chemtech MPS 350
1 Advanced Chemtech Apex 396
Instruments used in Protein Sequencing:
1 ABI Procise
1 ABI 477
1 ABI 420
Instruments used in Robotics:
1 Beckman Biomek 2000
1 Qiagen Biorobot 3000
Other advanced equipments used in genetics:
DMX 600: The instrument can be used with either a 1H/13C/15N triple resonance probe fitted with triple axis gradients or with a 1H/13C/ broadband X triple resonance probe.
DMX 500: The instrument can be configured to give optimum performance in the solution state for 1H, 13C or broadband multinuclear acquisitions or to perform 1H, 31P and broadband double and triple resonance experiments with z-gradients.
Varian VI 300: This spectrometer is a dedicated solid-state instrument with multinuclear, magic angle spinning and high rf power capabilities. 4mm and 7mm broadband probes are available.
Bruker DPX 300: This instrument is used largely for 2D structural studies and multinuclear solution measurements.
Bruker DPX 300: The instrument is fitted with a 60-holder sample changer and a four nuclei (QNP) z-gradient probe.