Characterization of a Microdissection Library from Human Chromosome Region 3p14

Assignment<footref rid="foot01">¹</footref> of human filamin gene FLNB to human chromosome band 3p14.3 and identification of YACs containing the complete FLNB transcribed region

“…Both FLNB primer pairs amplify products of the same length using cDNA or genomic DNA as templates. PCR analysis allowed us to define the STS-content of YACs from a previously established contig for chromosome region 3p14 (Bardenheuer et al, 1994;Michaelis et al, 1995) and to construct a contig containing the entire FLNB coding region and neighboring regions (Fig. 2).…”

“…Fluorescence in situ hybridization (FISH) and PCR analysis were performed as described earlier (Bardenheuer et al, 1994;Michaelis et al, 1995). Fig.…”

Identification of novel 'expressed sequence tags' within the FHIT gene locus in human chromosome region 3p14.2

“…Preparation of YAC DNA Yeast DNA containing YAC 268F1 (380 kb) was prepared as described previously (Bardenheuer et al 1994;Michaelis et al 1995). MboI digestion of YAC DNA (100 ng), ligation with 50 pmoles of MboI linkers, and amplification by polymerase chain reaction (PCR) using biotinylated oligonucleotides was performed as described elsewhere (Morgan et al 1992).…”

“…Reverse transcription and colony blot filter hybridizations were done as described (Sambrook et al 1989;Bardenheuer et al 1994). Filters were washed twice for 20 min in 0.1 × SSC, 0.1% SDS, and exposed to Kodak XAR films using intensifying screens for 1-5 days at -70°C.…”

Reassignment of EST w23312 to human chromosome 2q35→q37 by fluorescence in situ hybridization

“…Human chromosome region 3p14 has been proposed to harbor tumor suppressor genes involved in the development of lung cancer, renal cell carcinoma and other neoplasias. Lux et al (1997) identified a novel EST w23312 (D3S3862) from YAC clone 268F1 (380 kb) which had been mapped to 3p14 (Bardenheuer et al, 1994). The YAC was analyzed for its chimeric status by FISH (Michaelis et al, 1995).…”