Multi-Detection by Target Mixed Loop-Mediated Isothermal Amplification.

Loop-mediated isothermal amplification (LAMP), a nucleic acid amplification technology, has been characterized as having a short reaction time while generating a large amount of DNA product under isothermal conditions. Here, we found that LAMP is able to amplify more than one target simultaneously by using a mixture of human immunodeficiency virus- 1, hepatitis B virus, and hepatitis C virus primers, a process we have named target mixed LAMP. Target mixed LAMP and detection can be performed in less than 1 h. In this study, we also used target mixed LAMP to successfully amplify GAPDH, gamma-actin, TATA box binding protein, and haptoglobin from pooled cDNA derived from mouse tissues. Furthermore, using eight plasmid vectors as templates, target mixed LAMP is able to amplify all targets simultaneously. These findings reveal the usefulness of LAMP for the diagnosis of infectious disease as well as for conducting gene expression analysis.

DNA extraction from crayfish exoskeleton.

Crayfish exoskeleton (CE) samples are generally less invasive and easy to be collected. However, it is difficult to extract DNA from them. This study was intended to investigate CE as a DNA source and design an easy and efficient DNA extraction protocol for polymerase chain reactions. Specific primer pair (PPO-F, PPO-R) was used to amplify extracted DNA from CE, and compared to crayfish tail muscle DNA sample. Moreover, seven microsatellites markers were used to amplify the CE DNA samples set. Since the extracted DNA from CE is suitable for gene amplification, the results present usefulness of CE as an easy and convenient DNA source for PCR-based population genetic research.

Expression of Cellular Oncogenes in Gastric Carcinoma Related with Its Histologic Subtype: Southern blot analysis and immunohistochemistry

To search biologic characteristics of gastric carcinoma, one of the most common cancer in Korea, the author examined the alterations in DNA level and the expression of Ha-ras gene and c-myc gene in 20 primary tumors. Amplification of c-Ha-ras DNA was detected in 4(40%) of 10 patients who showed histologic subtype of relatively differentiated adenocarcinoma, but rearrangement of c-Ha-ras DNA was absent. Neither augumentation nor deletion of the c-myc DNA was observed. Higher expression of the ras p21 in tumor cells was noted in more differentiated tumor cells rather than poorly differentiated cases. One mucinous carcinoma, two signet ring cell carcinomas and one papillary carcinoma did not disclose expression of p21. The expressions of c-myc oncogene product were variable and were not correspond to the expressions of ras p21. A tendency that poorly differentiated tumor cells had higher expression of c-myc oncogene was suggested.

THE APPLICATION OF DNA AMPLIFICATION TECHNIQUE IN VITRO IN FORENSIC MEDICNE PRACTICE / 中国法医学杂志

This paper reports the determination of sex of human blood stains in threecriminal cases by the DNA amplification technique.The results fit the cases de-tails and gave the scientific evidence for the justice.The blood stains were dige-sted with proteinase K.The protein was extracted with the phenol-chloroform.The DNA was precipitated with NaCl and ethyl alcohol.Amplification of DNAwas carried out using two pairs of primer Y1.1 Y1.2 and Alu9.1 Alug.2 and heatstable FD polymerase.The amplified products were subjected to agarose gel(con-taining ethidium bromide)electrophoresis.Sex of blood stains was determinedaccording to the amplified products.PCR technique is rapid to perform,sensitiveand simple.No special equipments and isotope labelled probe are required.