[Construction and identification of the chromosome specific DNA library.].
The objective of the present study was to construct a human chromosome 21 specific DNA library for further use in research of genetic disease. Human chromosome 21 microdissected from the peripheral blood cells were subjected to repeatedly incubation in gradient temperature bath to release DNA. The library of chromosome 21 was constructed using the DNA fragment of 100-500 bp and 500-2 000 bp recovered from the products of DOP-PCR. Florescence in situ hy-bridization (FISH) and dot blotting analyses were carried out to assess the chromosome 21 specificity of the DNA library. The results indicated that DNA of chromosome 21 was released easily after repeatedly incubation in gradient temperature bath. Recovery of DNA fragments from DOP-PCR in different size ranges improved the efficiency of cloning of large fragments. Both FISH and dot blotting analyses revealed that the DNA library constructed in this study was chromosome 21-specific. This DNA library facilitates identification and investigation of the chromosome 21 related abnormality.
Sun
Lei
L
Qinzhou Women and Children Healthcare Hospital, Qinzhou, China. sunshijie12345@163.com
Liu
Xin-Xiong
XX
Chen
Zhe
Z
Zhang
Ming
M
Lu
Yang-Qing
YQ
Zhuo
Yong-Guang
YG
Fu
Ke-Peng
KP
Fan
Zu-Qian
ZQ
D002891:Chromosomes, Human, Pair 21; D015723:Gene Library; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016133:Polymerase Chain Reaction