Article archive
History.. how it all started and what it is all about..
30/09/2013 10:57
Recombinant DNA technology is the use of in vitro molecular techniques to isolate and manipulate fragments of DNA.
In the early 1970s, the first successes in making recombinant DNA molecules were accomplished Stanley Cohen at Stanford University and Herbert Boyer at the University...
A brief overview
15/09/2013 23:55An introduction...Cell based DNA cloning
11/09/2013 21:11
The fundamentals of current DNA technology are very largely based on two quite different approaches to studying specific DNA sequences within a complex DNA population
DNA cloning. The desired fragment must be selectively amplified so that it...
DNA Ligation(vector and insert)
11/09/2013 20:27
Ligation
The next step is the ligation of the insert into the linearized vector.
This involves the formation of phosphodiester bonds between adjacent 5'-phosphate and 3'-hydroxyl residues, which can be catalyzed by two different ligases: E. coli DNA ligase and bacteriophage T4 DNA...
Purification of DNA from living cells
11/09/2013 17:29
The genetic engineer will, at different times, need to prepare at least three distinct kinds of DNA.
Firstly, total cell DNA will often be required as a source of material from which to obtain genes to be cloned.
Total cell DNA may be DNA from a culture of bacteria, from a...
Transfer of DNA into host cells- the various methods
10/09/2013 21:35
The recombinant vector carrying foreign DNA needs to be transferred into the suitable host cells. Several methods have been developed for introduction of recombinant DNA molecule into host cells.
According to types of vectors and host cells, the methods are adopted. Some of the methods of gene...
Polymerase Chain Reaction
10/09/2013 20:36
PCR (Polymerase Chain Reaction) is a revolutionary method developed by Kary Mullis in the 1980s. PCR is based on using the ability of DNA Polymerase to synthesize new strand of DNA complementary to the offered template strand. Because DNA polymerase can add a nucleotide only...
Total Cell DNA
10/09/2013 17:56
Preparation of total cell DNA
The fundamentals of DNA preparation are most easily understood by first considering the simplest type of DNA purification procedure, that wbere the entire DNA complement of a bacterial cell is required. The modifications needed for plasmid and phage DNA...
Plasmid DNA
10/09/2013 16:23
Preparation of plasmid DNA
Purification of plasmids from a culture of bacteria involves the same general strategy as preparation of total cell DNA.
A culture of cells, containing plasmids, is grown in liquid medium, harvested, and a cell extract prepared. The protein and RNA are...
Phage DNA
10/09/2013 16:06
Preparation of bacteriophage DNA
The key difference between phage DNA purification and the preparation of either total cell DNA or plasmid DNA is that for phages the starting material is not normally a cell extract. This is because bacteriophage particles can be obtained in large...
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