PCR-quality DNA isolation from human bronchial aspirates and buccal and eyelid swabs by a simple procedure based on alkaline lysis.
There are only a few systematic reports about DNA extraction from routine diagnostic cytological specimens. An inevitable drawback of such techniques is increased spending of time and funds required for obligatory DNA purification. To implement a simple protocol for human DNA isolation from cytological specimens related to lung cancer, bronchial aspirates together with samples collected by swabbing of the inner cheek and eyelid were used. By combining alkaline and temperature lyses it was possible to isolate DNA solution ready for PCR in less than an hour. Testing the method used for amplification of sex chromatin gene fragments showed that it is highly efficient. The presented protocol preserves high-quality DNA that is suitable for PCR-based assays.
Vrabec Branica
Bozica
B
Department of Cytology, University Hospital for Lung Diseases Jordanovac, Zagreb, Croatia.
Mitrecić
Dinko
D
Grgić
Sandra
S
Smojver Jezek
Silvana
S
Chalfe
Nabil
N
Gajović
Srećko
S
D000328:Adult; D001980:Bronchi / Q000737:chemistry*; D002610:Cheek; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D005143:Eyelids / Q000737:chemistry*; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D012728:Sex Chromatin / Q000235:genetics; D013048:Specimen Handling / Q000379:methods*; D013396:Suction
D004247:DNA