Preparation of DNA Ladder Based on Multiplex PCR Technique

DNA molecular weight standard control, also called DNA marker (ladder), has been widely used in the experiments of molecular biology. In the paper, we report a method by which DNA marker was prepared based on multiple PCR technique. 100–1000 bp DNA fragments were amplified using the primers designed according to the 6631 ~ 7630 position of lambda DNA. Target DNA fragments were amplified using Touchdown PCR combined with hot start PCR, respectively, followed extracted by phenol/chloroform, precipitated with ethanol and mixed thoroughly. The results showed that the 100–1000 bp DNA fragments were successfully obtained in one PCR reaction, the bands of prepared DNA marker were clear, the size was right and could be used as control in the molecular biology experiment. This method could save time and be more inexpensive, rapid, simple when compared with the current DNA Ladder prepared means

Simple, Time-Saving Dye Staining of Proteins for Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis Using Coomassie Blue

A fixation-free and fast protein-staining method for sodium dodecyl sulfate–polyacrylamide gel electrophoresis using Coomassie blue is described. The protocol comprises staining and quick washing steps, which can be completed in 0.5 h. It has a sensitivity of 10 ng, comparable with that of conventional Coomassie Brilliant Blue G staining with phosphoric acid in the staining solution. In addition, the dye stain does not contain any amount of acid and methanol, such as phosphoric acid. Considering the speed, simplicity, and low cost, the dye stain may be of more practical value than other dye-based protein stains in routine proteomic research

Image processing based quantitative damage evaluation in composites with long pulse thermography

Pulsed thermography is a contactless and rapid non-destructive evaluation (NDE) technique that is widely used for the inspection of fibre reinforced plastic composites. However, pulsed thermography uses expensive and specialist equipment such high-energy flash lamps to generate heat into the sample, so that alternative thermal stimulation sources are needed. Long pulse thermography was recently developed as a cost-effective solution to enhance the defect detectability in composites by generating step-pulse heat into the test sample with inexpensive quartz halogen lamps and measuring the thermal response during the material cooling down. This paper provides a quantitative comparison of long pulse thermography with traditional pulsed thermography and step heating thermography in carbon fibre and glass fibre composites with flat-bottomed holes located at various depths. The three thermographic methods are processed with advanced thermal image algorithms such as absolute thermal contrast, thermographic signal reconstruction, phase Fourier analysis and principal component analysis in order to reduce thermal image artefacts. Experimental tests have shown that principal component analysis applied to long pulse thermography provides accurate imaging results over traditional pulsed thermography and step heating thermography. Hence, this inspection technique can be considered as an efficient and cost-effective thermographic method for low thermal conductivity and low thermal response rate materials. This work is carried out within the scope of EU H2020 funded EXTREME projec

1965 Volume 14 No. 2

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