Column spin type product for extracting total RNA from plant specimens such as leaves, seed, root, stem including a leguminous plant
• Possible to extract total RNA from various plant specimen
• Total RNA extraction is possible from seed samples containing large amounts of metabolites
– RNA extraction from fauna and legumes
– High purity and high yield of RNA extraction
– Possible to remove gDNA quickly and extract RNA within 20 minutes
– 99% removal of gDNA without DNase
The IQeasy™ Plus Plant RNA Extraction Mini Kit is designed to extract total RNA from tissue samples such as leaves, stems, roots, and seeds of various plant species.In plants, photosynthesis accumulates various metabolites and byproducts such as inhibitors. In order to remove inhibitors, Normally phenol use in the lysis step. However, in case where phenol is not used a high concentration of chaotrophic salt is used for lysis. In such cases, rice, barley, wheat, Beans and seeds (containing large amounts of protein and lipids), such as seeds (containing large amounts of starch) or soybeans, mung beans, peas, etc. In this case, the nucleic acid to be extracted is also precipitated (Co-precipitation) and the RNA content in the final recovered fraction is extremely low. IQeasy™ Plus Plant RNA Extraction Mini Kit is possible to overcome this case.
IQeasy™ Plus Plant RNA Extraction Mini Kit does not use chaotrophic salt during the lysis process to effectively extract total RNA from plant metabolites. Therefore, inhibitor is not eluted with RNA. It is designed to rapidly recover supernatant from homogenate using sieve spin column. It is designed to remove gDNA by applying 2 column system using gDNA binding column to minimize gDNA contamination. As mentioned earlier, there is no enzymatic lysis, therefore, It is possible to extract RNA easily within 20 minutes and it is possible to extract total RNA of high purity and high efficiency with one kit by providing two protocols which are divided into tissue and species.
IQeasy™ plus Series The Total RNA Extraction Mini Kit is an RNA extraction product (see Figure 1) that can effectively extract total RNA from a variety of samples. We provide specialized protocol including the lysis buffers and inhibitors removed for each sample, and minimize the gDNA content. Therefore, IQeasy™ Plus Series Total RNA Extraction Mini Kit is designed to be able to extract reliable high yield and high purity of total RNA. In addition, it is developed to be reproducible by providing specific protocol for each sample. All products of IQeasy™ Plus Series Total RNA Extraction Mini Kit can be used for pathogen research by using CAPS. We have significantly reduced concerns about contamination (see Figure 2). Therefore, RNA extracted using the IQeasy™ Plus Series Total RNA Extraction Mini Kit is a specialized product that can be applied to microarrays as well as pathogen research.
Figure 1. Four kinds of IQeasy™ plus Series Total RNA Extraction Mini Kits
Figure 2. Spin Column & CAPS Information
Recovery rate of total RNA from various plant species
Purity and concentration of total RNA extracted from various plant tissue using IQeasy™ Plus Plant RNA Extraction Mini Kit was identified. High purity of total RNA can be extracted from species that are difficult to extract due to secondary metabolites. High purity of total RNA can also be purified from seed samples containing high metabolites.
Analysis for quality of extracted RNA
Using IQeasy™RNA Plant Mini Kit, phenol-based RNA extraction kit and other RNA extraction kits, total RNA was extracted. After analyzing each extracted RNA quality with Agilent® 2100 bioanalyzer, extracted RNA using IQeasy™RNA Plant Mini Kit was verified that it was high yield and high purified RNA for Microarray analysis.
Lane M, RNA Ladder Marker; lane C, Positive control; lane 1, Leaf RNA extraction using easy-spin™ IIp Kit(Cat. No. 17310, iNtRON); lane 2, Leaf RNA extraction using Competitor’ kit; lane 3, Leaf RNA extraction using IQeasy™ Plus Plant Mini Kit; lane 4, Seed RNA extraction using Competitor’ kit; lane 5, Seed RNA extraction using IQeasy™ Plus Plant Mini Kit
Analysis for quantification through Real time PCR
After performing 1/10 serial dilution of extracted total RNA using IQeasy™ Plus Plant RNA Extraction Mini Kit, ONE-STEP RT-PCR PreMix Kit(iNtRON, Cat.No. 25101) was used to see efficiency of extraction by testing quantification for GAPDH Gene. Real-time RT-PCR was performed by high purified RNA.