Rapid zygosity determination in mice by SYBR Green real-time genomic PCR of a crude DNA solution

Transgenic Research

Volumn 17, Issue 1, 2008, Pages 149-155

  Sakurai, Takayuki ^a   Kamiyoshi, Akiko ^a   Watanabe, Satoshi ^b   Sato, Masahiro ^c   Shindo, Takayuki ^a  

^a SHINSHU UNIVERSITY GRADUATE SCHOOL OF MEDICINE   (Japan)

^b NATIONAL INSTITUTE OF AGROBIOLOGICAL SCIENCES   (Japan)

^c KAGOSHIMA UNIVERSITY   (Japan)

Author keywords

Crude DNA solution; Gene targeted mice; Real time PCR; SYBR Green; Transgenic mice; Zygosity

Indexed keywords

BETA GALACTOSIDASE; FLUORESCENT DYE; GTROSA26 PROTEIN, MOUSE; MEMBRANE PROTEIN; ORGANIC COMPOUND; PRIMER DNA; PROTEIN; RECEPTOR ACTIVITY MODIFYING PROTEIN; RECOMBINANT DNA; SIGNAL PEPTIDE; SYBR GREEN I; UNCLASSIFIED DRUG;

ANIMAL; ARTICLE; FEMALE; GENETICS; HETEROZYGOTE; HOMOZYGOTE; ISOLATION AND PURIFICATION; MALE; METHODOLOGY; MOUSE; MOUSE MUTANT; NUCLEOTIDE SEQUENCE; POLYMERASE CHAIN REACTION; PREGNANCY; TRANSGENIC MOUSE;

ANIMALS; BASE SEQUENCE; BETA-GALACTOSIDASE; DNA PRIMERS; DNA, RECOMBINANT; FEMALE; FLUORESCENT DYES; HETEROZYGOTE; HOMOZYGOTE; INTRACELLULAR SIGNALING PEPTIDES AND PROTEINS; MALE; MEMBRANE PROTEINS; MICE; MICE, KNOCKOUT; MICE, TRANSGENIC; ORGANIC CHEMICALS; POLYMERASE CHAIN REACTION; PREGNANCY; PROTEINS;

MUS; MUS MUSCULUS;

EID: 37549057664     PISSN: 09628819     EISSN: None     Source Type: Journal    
DOI: 10.1007/s11248-007-9134-7     Document Type: Article

References (12)

1. Bubner B, Gase KT, Baldwin IT (2004) Two-fold differences are the detection limit for determining transgene copy numbers in plants by real-time PCR. BMC Biotechnol 4:14
2. de Vries WN, Binns LT, Fancher KS, Dean J, Moore R, Kemler R, Knowles BB (2000) Expression of Cre recombinase in mouse oocytes: a means to study maternal effect genes. Genesis 26:110-112
3. Haurogné K, Bach JM, Lieubeau B (2007) Easy and rapid method of zygosity determination in transgenic mice by SYBR Green real-time quantitative PCR with a simple data analysis. Transgenic Res 16:127-131
4. Ji W, Zhou W, Abruzzese R, Guo W, Blake A, Davis S, Davis S, Polejaeva I (2005) A method for determining zygosity of transgenic zebrafish by TaqMan real-time PCR. Anal Biochem 344:240-246
5. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method 25:402-408
6. Mitrečić D, Huzak M, Ćurlin M, Gajović S (2005) An improved method for determination of gene copy numbers in transgenic mice by serial dilution curves obtained by real-time quantitative PCR assay. J Biochem Biophys Methods 64:83-98
7. Prior FA, Tackaberry ES, Aubin RA, Casley WL (2006) Accurate determination of zygosity in transgenic rice by real-time PCR does not require standard curves or efficiency correction. Transgenic Res 15:261-265
8. Sakurai T, Iguchi T, Moriwaki K, Noguchi M (1995) The ter mutation first causes primordial germ cell deficiency in ter/ter mouse embryos at 8 days of gestation. Dev Growth Differ 37:293-302
9. Shindo T, Kurihara H, Maemura K, Kurihara Y, Kuwaki T, Izumida T, Minamino N, Ju KH, Morita H, Oh-hashi Y, Kumada M, Kangawa K, Nagai R, Yazaki Y (2000) Hypotension and resistance to lipopolysaccharide-induced shock in transgenic mice overexpressing adrenomedullin in their vasculature. Circulation 101:2309-2316
10. Shitara H, Sato A, Hayashi J, Mizushima N, Yonekawa H, Taya C (2004) Simple method of zygosity identification in transgenic mice by real-time quantitative PCR. Transgenic Res 13:191-194
11. Tesson L, Heslan JM, Ménoret S, Anegon I (2002) Rapid and accurate determination of zygosity in transgenic animals by real-time quantitative PCR. Transgenic Res 11:43-48
12. Zambrowicz BP, Imamoto A, Fiering S, Herzenberg LA, Kerr WG, Soriano P (1997) Disruption of overlapping transcripts in the ROSA beta geo 26 gene trap strain leads to widespread expression of beta-galactosidase in mouse embryos and hematopoietic cells. Proc Natl Acad Sci USA 94:3789-3794