PMID- 15198206
OWN - NLM
STAT- MEDLINE
DCOM- 20050125
LR  - 20191108
IS  - 0962-8819 (Print)
IS  - 0962-8819 (Linking)
VI  - 13
IP  - 2
DP  - 2004 Apr
TI  - Simple method of zygosity identification in transgenic mice by real-time 
      quantitative PCR.
PG  - 191-4
AB  - To determine zygosity in transgenic (Tg) mice, a new technology, real-time 
      quantitative PCR, has recently been introduced in transgenic research to overcome 
      several drawbacks (time-consuming, specialized techniques and/or ambiguity in the 
      results) of previously established methods, for example, Southern blot 
      hybridization, dot blot hybridization, fluorescence in situ hybridization (FISH), 
      etc. However, the previous real-time quantitative PCR method still possesses 
      several drawbacks, for example, it needs two sets of primers/probes and the 
      complicated setting up of appropriate conditions, both of which are expensive and 
      remain time-consuming. We therefore developed an improved real-time quantitative 
      PCR system for determination of zygosity, which is easy, rapid and less 
      expensive, because the technique needs only two experimental processes: 
      estimation of DNA concentration and CYBR Green PCR. We found that homozygous, 
      hemizygous and non-Tg animals could easily be distinguished among F1 littermates 
      in crosses of hemizygous EGFP- and DsRed2-Tg mice. Our improved method will be 
      applicable to any Tg mouse strains, when a primer set is matched to the 
      corresponding transgene.
FAU - Shitara, Hiroshi
AU  - Shitara H
AD  - Laboratory for Transgenic Technology, The Tokyo Metropolitan Institute of Medical 
      Science, Tokyo 113-8613, Japan. shitara@rinshoken.or.jp
FAU - Sato, Akitsugu
AU  - Sato A
FAU - Hayashi, Jun-ichi
AU  - Hayashi J
FAU - Mizushima, Noboru
AU  - Mizushima N
FAU - Yonekawa, Hiromichi
AU  - Yonekawa H
FAU - Taya, Choji
AU  - Taya C
LA  - eng
SI  - GENBANK/U15541
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Transgenic Res
JT  - Transgenic research
JID - 9209120
RN  - 0 (Fluorescent Dyes)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Fluorescent Dyes/analysis
MH  - Genetic Carrier Screening/*methods
MH  - *Heterozygote
MH  - *Homozygote
MH  - Mice
MH  - Mice, Transgenic/*genetics
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods
EDAT- 2004/06/17 05:00
MHDA- 2005/01/26 09:00
CRDT- 2004/06/17 05:00
PHST- 2004/06/17 05:00 [pubmed]
PHST- 2005/01/26 09:00 [medline]
PHST- 2004/06/17 05:00 [entrez]
AID - 10.1023/b:trag.0000026084.32492.eb [doi]
PST - ppublish
SO  - Transgenic Res. 2004 Apr;13(2):191-4. doi: 10.1023/b:trag.0000026084.32492.eb.