TY - JOUR
T1 - CIS - Cloning of identical sequences between two complex genomes
AU - Zabarovska, Veronika
AU - Li, Jingfeng
AU - Muravenko, Olga
AU - Fedorova, Ludmila
AU - Braga, Eleonora
AU - Ernberg, Ingemar
AU - Wahlestedt, Claes
AU - Klein, George
AU - Zabarovsky, Eugene R.
N1 - Funding Information:
This work was supported by research grants from Swedish Cancer Society, Pharmacia & Upjohn, Kar-olinska Institute, Ingabritt och Arne Lundbergs For-skningsstiftelse and Åke Wiberg foundation. VZ was a recipient of a fellowship from the Concern Foundation in Los Angeles and the Cancer Research Institute in New York. OM was a recipient of a fellowship from the Swedish Institute.
PY - 2000
Y1 - 2000
N2 - Development of the methods permitting cloning of identical sequences between two sources of DNA can be very useful for many purposes, including isolation of disease genes. Here we describe a new method called CIS (cloning of identical sequences). A combination of digestion with MvnI, treatment with mung bean nuclease, UDG (uracil-DNA glycosylase) and PCR with 5'-methyl-dCTP and dUTP was used to isolate identical sequences between two micro-cell hybrid lines (MCH). In a control experiment, mouse MCH903.1 and MCH939.2 containing human chromosome 3 from different individuals, were compared using the CIS procedure. Only background fluorescence in-situ hybridization (FISH) was achieved. In another experiment, mouse MCH903.1, containing complete human chromosome 3, and rat MCH429.11, containing a part of human 3q from the same chromosome were compared. The experiment showed that the original MCH429.11 and the DNA purified using the CIS procedure had identical FISH patterns to human metaphase chromosomes, thus demonstrating the efficiency of CIS.
AB - Development of the methods permitting cloning of identical sequences between two sources of DNA can be very useful for many purposes, including isolation of disease genes. Here we describe a new method called CIS (cloning of identical sequences). A combination of digestion with MvnI, treatment with mung bean nuclease, UDG (uracil-DNA glycosylase) and PCR with 5'-methyl-dCTP and dUTP was used to isolate identical sequences between two micro-cell hybrid lines (MCH). In a control experiment, mouse MCH903.1 and MCH939.2 containing human chromosome 3 from different individuals, were compared using the CIS procedure. Only background fluorescence in-situ hybridization (FISH) was achieved. In another experiment, mouse MCH903.1, containing complete human chromosome 3, and rat MCH429.11, containing a part of human 3q from the same chromosome were compared. The experiment showed that the original MCH429.11 and the DNA purified using the CIS procedure had identical FISH patterns to human metaphase chromosomes, thus demonstrating the efficiency of CIS.
KW - Chromosome mapping
KW - Genetic technique
KW - Human genetics
KW - In-situ hybridization methods
KW - Molecular cloning
UR - https://www.scopus.com/pages/publications/0034011771
U2 - 10.1023/A:1009243606611
DO - 10.1023/A:1009243606611
M3 - Article
C2 - 10730592
AN - SCOPUS:0034011771
SN - 0967-3849
VL - 8
SP - 77
EP - 84
JO - Chromosome Research
JF - Chromosome Research
IS - 1
ER -