Chapter 24
Fluorescence in Situ Hybridisation of a Complete Set of
41 Telomere Specific Probes in a Single Simultaneous
Hybridisation Experiment Using the Chromoprobe
Multiprobe System-T
MARTIN LAWRIE, MARIA OCANA GIL, and DON CARDY
Introduction
It is estimated that around 3% of the population exhibit some degree of
mental retardation where the pathogenesis of their disorder is poorly understood (1). The importance of subtelomeric chromosome rearrangements associated with this idiopathic mental retardation (IMR) has
been shown to be as much as 8% in moderate to severe retardation
and of 0.5% in moderate retardation (2, 3, 4). There are also reports of
disruption of these regions of the chromosomes being implicated in recurrent miscarriages, haematological malignancies and polycystic kidney
disease (5).
The detection of such subtle chromosome rearrangements in patients
suspected ofhaving a chromosome anomaly has up to now been limited to
standard cytogenetic staining techniques with a limit of 850-1000 band
resolution. This is not only time-consuming but also limited because
of the similar banding patterns (G negative) found at the telomeres.
Thus, cryptic deletions and rearrangements can prove to be impossible
to detect by conventional methods.
The advent of Fluorescence in situ hybridisation (FISH) technology has
made new approaches available for cytogenetic screening (6,7). Whole
chromosome painting and reverse chromosome paints have not, however,
been able to address the screening needs of the telomeres because these
regions are underrepresented in the whole chromosome libraries so that
~ Martin Lawrie, Cytocell Limited, Somerville Court, Banbury Business Park,
Adderbury, Banbury, Oxfordshire, OX17 3SN, UK (phone +44-1295-810910;
fax +44-1295-812333; e-mail m_lawrie@cytocell.co.uk)
Maria Ocana Gil, Cytocell Limited, Somerville Court, Banbury Business Park,
Adderbury, Banbury, Oxfordshire, OX17 3SN, UK
Don Cardy, Cytocell Limited, Somerville Court, Banbury Business Park, Adderbury,
Banbury, Oxfordshire, OX17 3SN, UK
PROTOCOL
Fluorescence in Situ Hybridisation of a Complete Set of
41 Telomere Specific Probes in a Single Simultaneous
Hybridisation Experiment Using the Chromoprobe
Multiprobe System-T
MARTIN LAWRIE, MARIA OCANA GIL, and DON CARDY
Introduction
It is estimated that around 3% of the population exhibit some degree of
mental retardation where the pathogenesis of their disorder is poorly understood (1). The importance of subtelomeric chromosome rearrangements associated with this idiopathic mental retardation (IMR) has
been shown to be as much as 8% in moderate to severe retardation
and of 0.5% in moderate retardation (2, 3, 4). There are also reports of
disruption of these regions of the chromosomes being implicated in recurrent miscarriages, haematological malignancies and polycystic kidney
disease (5).
The detection of such subtle chromosome rearrangements in patients
suspected ofhaving a chromosome anomaly has up to now been limited to
standard cytogenetic staining techniques with a limit of 850-1000 band
resolution. This is not only time-consuming but also limited because
of the similar banding patterns (G negative) found at the telomeres.
Thus, cryptic deletions and rearrangements can prove to be impossible
to detect by conventional methods.
The advent of Fluorescence in situ hybridisation (FISH) technology has
made new approaches available for cytogenetic screening (6,7). Whole
chromosome painting and reverse chromosome paints have not, however,
been able to address the screening needs of the telomeres because these
regions are underrepresented in the whole chromosome libraries so that
~ Martin Lawrie, Cytocell Limited, Somerville Court, Banbury Business Park,
Adderbury, Banbury, Oxfordshire, OX17 3SN, UK (phone +44-1295-810910;
fax +44-1295-812333; e-mail m_lawrie@cytocell.co.uk)
Maria Ocana Gil, Cytocell Limited, Somerville Court, Banbury Business Park,
Adderbury, Banbury, Oxfordshire, OX17 3SN, UK
Don Cardy, Cytocell Limited, Somerville Court, Banbury Business Park, Adderbury,
Banbury, Oxfordshire, OX17 3SN, UK
PROTOCOL