CIS--cloning of identical sequences between two complex genomes.
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.
Zabarovska
V
V
Microbiology & Tumor Biology Center (MTC), Karolinska Institute, Stockholm, Sweden.
Li
J
J
Muravenko
O
O
Fedorova
L
L
Braga
E
E
Ernberg
I
I
Wahlestedt
C
C
Klein
G
G
Zabarovsky
E R
ER
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D004247:DNA