Biomedical subjects

Hong-min Wang

Publications and source records attributed to Hong-min Wang.

5 recordsLinked to original sources

PubMed2005

[Comparison of two amine-modified chemical platforms for DNA microarray preparation].

OBJECTIVE: To study two amine modification procedures for DNA microarray preparation based on polymeric coatings. METHODS: One of the proposed approaches utilized poly-amine coating of silanized slides activated by 1,4-phenylene diisothiocyanate, and the other employed acrylic acid-co-acrylamide copolymer and 1-(3-dimethylamino propyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide as the coating agents and activator, respectively. The modified slides were used for preparing lambda phage DNA microarrays, whose properties were analyzed by hybridization. RESULTS: Formation of dendrimeric structure and polymer was observed on the surface of the slides. The signal spots in uniform, steady and regular shape, in comparison with the commercial CMT-GAPS slides, indicated successful manufacture of the microarrays. CONCLUSION: The two platforms are suitable for microarray preparation, and the method of acrylic acid-co-acrylamide copolymer modification is more preferred.

Acrylamides↗

PubMed2004

DNA microarray probe preparation by gel isolation nested PCR.

To develop a simplified method that can rapidly prepare DNA microarray probes in a massive scale, a lambda phage genomic DNA-fragments library was constructed for the microarray-probes collection. Four methods of DNA band recovery from the first PCR products were tested and compared. The DNA microarray probes were collected by a novel method of nested PCR that was mediated by gel isolation of the first PCR products. This method was named GIN-PCR. The probes that were prepared by this GIN-PCR technique were used as subjects to fabricate a DNA microarray. The results showed that a wooden toothpick was superior to the other 3 methods, since this technique can steadily transfer the DNA bands as the template of the second PCR after the first PCR. A group of probes were successfully collected and DNA microarrays were constructed using these probes. Hybridization results demonstrated that this technique of DNA recovery and probe preparation was rapid, efficient, and effective. We developed a cost-effective and less labor-intensive method for DNA microarray probe preparation by nested PCR that is mediated by wooden toothpick transfer of the DNA bands in the gel after electrophoresis.

Bacteriophage lambda↗

PubMed2004

Design and preparation of oligonucleotide microarray for vaccinia virus detection.

OBJECTIVE: To study the preparation of oligonucleotide microarray for detecting vaccinia virus. METHODS: Oligonucleotide probes were designed and synthesized according to the specific genes of vaccinia virus. Sample DNA of the virus and the negative control sample were obtained and labeled by restriction display technique, followed by hybridization to the oligonucleotide microarray and scanned by Agilent scanner. RESULTS: Strong hybridization signals were detected from the viral DNA hybridized with the microarray, but were absent in the negative sample when positive probes were not used. CONCLUSION: Distinct differences in the hybridization signals between the virus sample and negative sample and between the samples obtained in different phase of infection demonstrate high specificity and sensitivity of the microarray for vaccinia virus detection.

Base Sequence↗

PubMed2003

[Gene sequence analysis of SARS-associated coronavirus by nested RT-PCR].

OBJECTIVE: To explore an effective means for the detection of Severe Acute Respiratory Syndrome (SARS)- associated coronavirus. METHODS: The RNAs of the virus contained in the sputum samples from established SARS patients were extracted and reversely transcripted, followed by nested PCR using the reversely transcripted cDNA as the template. The PCR products were cloned then into the pMD18-T vectors, followed by sequence analysis. RESULTS: Specific fragments were amplified from the sputum samples of SARS patients, which were confirmed by DNA cloning and sequencing to belong to SARS-associated coronavirus. The Result of Blast shows only the difference in one nucleic acid from the TOR2 strain of SARS-associated coronavirus. CONCLUSION: Sequence analysis has confirmed the existence of SARS-associated coronavirus in the sputum samples of SARS patients, and nested RT-PCR is a quick, easy, and convenient way for the detection of the virus.

Base Sequence↗

PubMed2003

Quick preparations of human parvovirus B19 microarray probes using PCR.

OBJECTIVE: To prepare DNA microarray probes for the detection of human parvovirus B19. Method Specific PCR primers were designed with the Primer Premier 5.0 to amplify the conserved regions of human parvovirus B19 genome. The PCR products were cloned into the pMD-18 T vector. METHOD: Sequences analysis showed the PCR products conformed to the sequences contained in the genome of human parvovirus B19. RESULT: Sequences analysis showed the PCR products conformed to the sequences contained in the genome of human parvovirus B19. CONCLUSION: PCR amplification of the conserved and specific human parvovirus B19 genes is simple and effective to prepare the desired probes.

DNA Probes↗