9780521516358book.pdf

enzymes have been characterised that recognise over 100 different target sequences. A number of these, termedisoschizomers, reco ...

It is possible to check the integrity of the DNA byagarose gel electrophoresisand determine the concentration of the DNA by usin ...

be worn, and a strong detergent should be included in the isolation medium to immediately denature any RNases. Subsequent deprot ...

Contaminants may also be identified in the same way as that for DNA by scanning UV spectrophotometry; however, in the case of RN ...

5.7.3 Automated and kit-based extraction of nucleic acids Most of the current reagents used in molecular biology and the most co ...

DNA to migrate out of the gel piece, but it remains trapped within the dialysis tubing, and can therefore be recovered easily. 5 ...

to be captured within a very short timescale. More recently alternative methods of analysis including high performance liquid ch ...

regions. A number of bioinformatics resources such as GRAIL can be used to identify such features in a DNA sequence. 5.9 Molecul ...

as simple as this, and bioinformatic analysis of the restriction fragment lengths is usually needed to construct a map. 5.9.2 Nu ...

thestringencyof the hybridisation and it is particular for each individual gene probe and for each sample of DNA. A series of wa ...

point with which to produce a so-called heterologous gene probe. Although in some cases probes are already produced and cloned i ...

Perhaps the most common radioactive label is 32-phosphorus (^32 P), although for certain techniques 35-sulphur (^35 S) and triti ...

Using the other end of the DNA molecule, the 3^0 end, is slightly less complex. Here a new dNTP which is labelled (e.g.^32 P-adA ...

30 exonuclease activity. This is prepared by cleavage of DNA polymerase with subtilisin, giving a large enzyme fragment which ha ...

and a quencher molecule at the other. The oligonucleotide has a stem–loop structure where the stems place the fluorophore and qu ...

reaction and relative ease of the practical manipulation steps. Indeed combined with the relevant bioinformatics resources for i ...

analogous to the DNA replication process that takes place in cells since the outcome is the same: the generation of new compleme ...

amplified products at all. The annealing step allows the hybridisation of the two oligonucleotide primers, which are present in ...

Cycle 1 5  3  5  3  3  5  3  5  3  5  3  5  Cycle 2 5  3  3  5  Cycle 3 5  3  3 ^5  5  3  3  5  5  3  ...

arbitrary priming in genomic templates but interestingly may give rise to discrete banding patterns when analysed by gel electro ...