Isolation of nucleic acids from plant tissues using molecular kits
Plant DNA Isolation has unique challenges that require kits specifically designed to deal with carbohydrates, phenolics, and other compounds abundant in plant tissues.
Plant cell walls can be very difficult to disrupt, and lysates often contain significant amounts of compounds such as tannins, phenolics, and complex polysaccharides that can affect DNA and RNA quality and inhibit downstream reactions.
Life Technologies offers an expansive portfolio of kits, reagents, and devices for the isolation, purification, analysis, and quantitation not only for plant genomic DNA, but also for total RNA, mRNA, and microRNA or transcriptome RNA.
Which Plant DNA or RNA Isolation Kit is Right for You? Click here to find out
CCK-8 for Natural Products Analysis in Treatment of Cancer
Thousands of different species of plant have been reported as being useful for the treatment of cancer. There has been an argument that the use of natural products has been the single most successful strategy in the discovery of novel medicines. Tulp, Martin; Bohlin, Lars (2002)
Dojindo’s Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays of your natural product analysis.
Benefits of CCK-8:
- No Thawing Necessary- 3 simple steps
- Stable One Bottle Solution, at 4ºC for 1 year
- More Sensitive than MTT, MTS, or WST-1
- No Toxicity to Cell
Refer below for some publications related to CCK-8 used in natural products analysis for cancer treatment:
1. Extracts from Curcuma zedoaria Inhibit Proliferation of Human Breast Cancer Cell MDA-MB-231 In Vitro
This study was conducted to evaluate the effect of petroleum ether extracts of Curcuma zedoaria on the proliferation of human triple negative breast cancer cell line MDA-MB-231. The sample was assayed by CCK8 for MDA-MB-231 cellular viability with various concentrations and days before cell cycle analyses, Western Blot analysis, and Realtime Reverse Transcriptase PCR analyses for chemokines molecules were conducted. Read more
2. Sulforaphane induces cell cycle arrest by protecting RB-E2F-1 complex in epithelial ovarian cancer cells
Sulforaphane (SFN), an isothiocyanate phytochemical present predominantly in cruciferous vegetables such as brussels sprout and broccoli, is considered a promising chemo-preventive agent against cancer. This paper is a study conducted to understand the signaling mechanisms through which SFN influences the cell growth and proliferation in epithelial ovarian cancer cells (EOC). Cell proliferation assays were conducted using Cell Counting Kit-8 (CCK-8). Read more
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Purification of Phenolic Compound from Curcuma longa (Turmeric) Using Mass Directed Fractionation
The purification of natural products remains challenging, lengthy, and tedious. Automated flash chromatography instruments ease this process.
Reversed Phase Liquid Chromatography-Mass Spectrometry (LC-MS) has become a routine technique in today’s laboratory. A study involving an experimental Flash Chromatography-
Mass Spectrometry (FCMS) system, CombiFlash® Rf+ PurIon™ was used with both normal and reversed phase solvent systems to screen for novel compounds in Curcuma longa (turmeric) during purification.
Curcuma longa is
valued for the yellow color it imparts to food, and has also been
implicated in wound healing, anti-oxidant, anti-inflammatory,
anti-carcinogenic effects and natural coloring activities.
From this study, it has been proven that by using a mass spectrometer as a detector, potentially interesting compounds can be easily identified, prioritized, and purified in sufficient quantities to allow evaluation for biological activity.
The system can be run either in SIC mode to capture a particular compound in a mixture or in XIC to purify complex mixtures from natural products extracts. In either mode, the system collects the entire mass spectrum so that the other compounds can be identified..
Please click here to download the application note.
Optimized DNA Extraction from Basil lysed with Geno Grinder
Effective cell disruption and DNA/RNA extraction are critical for natural products research. SPEX SamplePrep’s Geno Grinder aids your research by providing high-throughput sample preparation solutions.
Geno Grinder functions as mechanical cell lysers used for effective extraction of genomic DNA from samples containing plant tissue. However, all mechanical lysers do not provide the same outcome.
A study was performed to discover whether Geno/Grinder processing variables significantly increased or decreased the yields of genomic DNA in fresh basil. Variables investigated included homogenization time and speed, while buffers, grinding media, and tubes were constant. In this experiment, the Geno Grinder returned most results higher than 6,557bp meaning that significant shearing or other degradation did not occur even with longer processing times.
The 2000 Geno/Grinder® was compared with a competitive cell lyser (Competitor A) for extraction of DNA from fresh basil. Results indicated that use of the Geno/Grinder provided DNA with higher molecular weight than the competitive instrument. In addition, optimal homogenization conditions using the Geno/Grinder were found to be 90 sec. at a rate of 2000 cyc/min.
Download the application note.