Optimization and evaluation of single-cell whole, genome multiple displacement amplification

被引：125

作者：

Spits, C ^{[1
]}

Le Caignec, C ^{[1
]}

De Rycke, M ^{[1
]}

Van Haute, L ^{[1
]}

Van Steirteghem, A ^{[1
]}

Liebaers, I ^{[1
]}

Sermon, K ^{[1
]}

机构：

[1] Vrije Univ Brussels, Acad Ziekenhuis, Res Ctr Reprod & Genet, B-1090 Brussels, Belgium

来源：

HUMAN MUTATION | 2006年 / 27卷 / 05期

关键词：

WGA; MDA; single cell; Phi29 DNA polymerase; Bst DNA polymerase;

D O I：

10.1002/humu.20324

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The scarcity of genomic DNA can be a limiting factor in some fields of genetic research. One of the methods developed to overcome this difficulty is whole genome amplification (WGA). Recently, multiple displacement amplification (MDA) has proved very efficient in the WGA of small DNA samples and pools of cells, the reaction being catalyzed by the 29 or the Bst DNA polymerases. The aim of the present study was to develop a reliable, efficient, and fast protocol for MDA at the single-cell level. We first compared the efficiency of 29 and Bst polymerases on DNA samples and single cells. The 29 polymerase generated accurately, in a short time and from a single cell, sufficient DNA for a large set of tests, whereas the Bst enzyme showed a low efficiency and a high error rate. A single-cell protocol was optimized using the phi 29 polymerase and was evaluated on 60 single cells; the DNA obtained DNA was assessed by 22 locus-specific PCRs. This new protocol can be useful for many applications involving minute quantities of starting material, such as forensic DNA analysis, prenatal and preimplantation genetic diagnosis, or cancer research.

引用

页码：496 / 503

页数：8

共 27 条

[1] Comparison of yield and genotyping performance of multiple displacement OmniPlex™ whole genome amplified DNA generated from multiple DNA sources [J].

Bergen, AW ;

Haque, KA ;

Qi, Y ;

Beerman, MB ;

Garcia-Closas, M ;

Rothman, N ;

Chanock, SJ .

HUMAN MUTATION, 2005, 26 (03) :262-270

[2]

BLANCO L, 1989, J BIOL CHEM, V264, P8935

[3] Whole genome amplification using a degenerate oligonucleotide primer allows hundreds of genotypes to be performed on less than one nanogram of genomic DNA [J].

Cheung, VG ;

Nelson, SF .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (25) :14676-14679

[4] Comprehensive human genome amplification using multiple displacement amplification [J].

Dean, FB ;

Hosono, S ;

Fang, LH ;

Wu, XH ;

Faruqi, AF ;

Bray-Ward, P ;

Sun, ZY ;

Zong, QL ;

Du, YF ;

Du, J ;

Driscoll, M ;

Song, WM ;

Kingsmore, SF ;

Egholm, M ;

Lasken, RS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (08) :5261-5266

[5] Rapid amplification of plasmid and phage DNA using phi29 DNA polymerase and multiply-primed rolling circle amplification [J].

Dean, FB ;

Nelson, JR ;

Giesler, TL ;

Lasken, RS .

GENOME RESEARCH, 2001, 11 (06) :1095-1099

[6]

Eckert K A, 1991, PCR Methods Appl, V1, P17

[7]

ESTEBAN JA, 1993, J BIOL CHEM, V268, P2719

[8] Improving clinical preimplantation genetic diagnosis for cystic fibrosis by duplex PCR using two polymorphic markers or one polymorphic marker in combination with the detection of the ΔF508 mutation [J].

Goossens, V ;

Sermon, K ;

Lissens, W ;

De Rycke, M ;

Saerens, B ;

De Vos, A ;

Henderix, P ;

Van de Velde, H ;

Platteau, P ;

Van Steirteghem, A ;

Devroey, P ;

Liebaers, I .

MOLECULAR HUMAN REPRODUCTION, 2003, 9 (09) :559-567

[9] Isothermal whole genome amplification from single and small numbers of cells: a new era for preimplantation genetic diagnosis of inherited disease [J].

Handyside, AH ;

Robinson, MD ;

Simpson, RJ ;

Omar, MB ;

Shaw, MA ;

Grudzinskas, JG ;

Rutherford, A .

MOLECULAR HUMAN REPRODUCTION, 2004, 10 (10) :767-772

[10] Clinical application of multiple displacement amplification in preimplantation genetic diagnosis [J].

Hellani, A ;

Coskun, S ;

Tbakhi, A ;

Al-Hassan, S .

REPRODUCTIVE BIOMEDICINE ONLINE, 2005, 10 (03) :376-380

← 1 2 3 →