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probes and primers are usually designed with bioinformatics software using sequence information from nucleic acid databases. Alt ...

superior to cloned double-stranded probes indicated in Section 6.4.1. They are used extensively inin situhybridisation and for i ...

Nylon membrane Master plate Grow bacterial colonies harbouring plasmid Colonies defined by autoradiography are used for further ...

screening may be undertaken in 3–4 h whereas it may be several days before detection by hybridisation is achieved. The PCR scree ...

Colonies from cDNA library on agar plate Isolate DNA from each clone, denature and bind to nylon membranes Add total mRNA under ...

hybrid arrest translation a positive result is indicated by the absence of a particular translation product when total mRNA is h ...

Recombinantλgt11 vector cDNA insert β-Galactosidase lacpromoter In vitro packaging Plate on bacterial lawn Induce production of ...

that makes M13 useful in chain termination sequencing is the placement of universal priming sites at20 or40 bases from the sta ...

contain a sequence with the mutation and half contain the wild-type sequence. Plaque hybridisation using the oligonucleotide as ...

from each reaction to hybridise. Following this, one of the two hybrids bearing a free 30 hydroxyl group is extended to produce ...

error-prone PCRwhere deliberate and random mutations are introduced by a low- fidelity PCR amplification reaction. The resulting ...

of foreign genes. In general the expression of foreign genes is carried out in specialised cloning vectors (Fig. 6.34). However ...

proteins may be detected by polyacrylamide gel electrophoresis or by immunological detection usingwestern blotting. Recently oli ...

sites for the insertion of foreign DNA. These regulatory sequences, such as that from the lacoperon ofE. coli, are usually deriv ...

in particular has been used to correct the defective ion transport by introducing a wild-type cystic fibrosis gene into cells be ...

linked to a major coat protein of a coliphage whilst the single-stranded gene encoding the protein is packaged within the virion ...

as well as naturally existing ones. This strategy is not restricted to antibodies and vast libraries of peptides may be used in ...

detection of particular RNA transcripts by blotting extracted mRNA and immobilising it to a nylon membrane (Section 5.9.2). Subs ...

detecting, for example, viral gene expression and furthermore allows the means of differentiating between latent and active viru ...

6.8.2 Analysing genesin situ Gross chromosomal changes are often detectable by microscopic examination of the chromosomes within ...