[Rapid detection of SOX2 gene by primed in situ labeling].
OBJECTIVE
To rapidly detect SOX2 gene using primed in situ labeling (PRINS).


METHODS
Human peripheral blood samples were cultured using an optimized method. Sequence of the SOX2 gene was amplified in situ with biotin-labeled specific primers and processed with a tyramide signal amplification (TSA) biotin system. Subsequently, fluorescence-stained signal was detected by streptavidin-Texas red. For the control group, MCF-10F cells were transfected with Lentivirus hSox2.


RESULTS
By VideoTesT-FISH software analysis, the long arm of chromosome 3 in the experimental group showed a specific red fluorescence signal, whilst the control samples showed no specific signals for SOX2. Transfected MCF-10F cells showed various efficiency of SOX2 gene integration.


CONCLUSIONS
PRINS utilizes a highly sensitive in situ PCR technique combined with fluorescence labeled oligodeoxynucleotides can synthesize probes in situ, thus greatly reducing the cost of probe and time for detection. It can facilitate identification and classification of induced pluripotent stem cells, and has many potential applications in this prospect.
Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Institute of Medical Genetics, College of Life Science, Sichuan University, Chengdu, Sichuan 610041, P. R. China.
Xi
X
Luo
Xianping
X
Ding
Lin
L
Chen
Qiang
Q
Quan
D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008297:Male; D020401:Primed In Situ Labeling / Q000379:methods*; D055748:SOXB1 Transcription Factors / Q000737:chemistry*
C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors