CONTROLLING SPEED WITH AE PIXEL SORTER 2 MANUAL
The yields for RNA and DNA are similar between manual and automation. The average yields are shown in figure 1. RNAĭNA and RNA was extracted from 3 10 µM curls of breast, intestine, and lung FFPE tissue. After the incubation for the DNase treatment, the user must add 150 µL of RBA the automated processing can proceed on from there. First the protocol is automated up to the DNase I treatment. The automation for RNA extraction is a two-step process. The optimized parameters and reagent volumes for RNA processing are in Table 1 and for DNA processing are in Table 2. The parameters for mix times and speeds and the collect times were optimized for the magnetic beads on the KingFisher™ Duo Prime. The plate was then moved to the KingFisher™ Duo Prime for DNA processing. The mixture was incubated at room temperature for 5 minutes. RNase A was added to the 100 µL lysate and mixed 10 times. After the incubation 100 µL was removed and added to a KingFisher™ Duo Prime 96 well plate. The remaining lysate was incubated at 60☌ for another hour followed by an 80☌ incubation for 1 hour. After the 2 hour incubation 100 µL of the lysate was removed and added to a KingFisher™ Duo Prime 96 well plate for RNA processing. The samples were then incubated at 60☌ for 2 hours. Following the incubation the tubes were allowed to cool for 5 minutes before the addition of 30 µL of proteinase K. The samples were incubated again at 80☌ for 5 minutes. Then 200 µL of LBD was added and the samples were centrifuged according to the protocol. The samples were then incubated at 80☌ for 5 minutes. For each tissue type 450 µL of mineral oil was added to each sample. 10 µM curls were cut and three samples consisting of 3 curls were processed both manually and on automation. In this study three FFPE tissue samples were used to assess the differences between manual and semi-automated RNA and DNA extraction. Automating the chemistry can also reduce the risk of human error, reduce hands-on time and total time therefore giving the user the ability to run more samples in a day. By automating FormaPure XL Total chemistry on the KingFisher™ Duo Prime the difficulty of working with FFPE tissue is eased. FormaPure XL Total has been demonstrated to obtain high quality FFPE-derived nucleic acids. This application note demonstrates the use of FormaPure XL Total, a total nucleic acid extraction kit for FFPE samples, in conjunction with the KingFisher™ Duo Prime Sample Purification System as a potential solution that mitigates some of the challenges with FFPE workflows. A solution that offers the ability to apply high-throughput and reliable genomic applications to FFPE samples can help accelerate cancer research and biomarker discovery. Second, consistent results for most high-throughput and automated extraction methods for FFPE are difficult to achieve due to inherent differences in the variety of tissue and disease types. First, FFPE samples generally provide low-quality nucleic acids, especially the RNA, as RNA undergoes more severe degradation and chemical and covalent modifications due to the effects of formalin-fixation than DNA. However, challenges exist in most FFPE workflows, especially when high-throughput extractions are a requirement. Current next-generation sequencing (NGS) technologies and extraction methods make it possible to study and identify cancer-causing alterations at the genomic and transcriptomic levels. Инструменты управления клинической информациейįormalin-fixed paraffin-embedded tissues are an invaluable resource for histological and genetic testing for cancer.
CONTROLLING SPEED WITH AE PIXEL SORTER 2 SERIES
CytoFLEX Violet-Blue-Red Series Upgrades.Navios EX – Powerful, Dependable Flow Cytometer.
0 kommentar(er)
