A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium
acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple
samples of organelle DNA from a small amount of tissue.
DNA Microprep Isolation from Plants Protocol The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of isolating DNA from plant material very rapidly. The procedure requires a table-top drill-press (mechanized homogenizer).
DNA Miniprep Isolation from Plants Protocol The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of speed and its use of inexpensive reagents.
Isolation of Retroelement from Plant Genomic DNA Retroelements and their derivatives are a ubiquitous and abundant component of plant genomes. Major classes of retroelements include the Pseudoviridae (Ty1-copia ), the Metaviridae (Ty3 -gypsy) and the Retroposineae LINE (non-LTR) groups. All reverse transcribing elements can be included in a universal classification. Includes: Pseudoviridae (Ty1-copia) Degenerate Primers; Metaviridae (Ty3-gypsy) Element Degenerate Primers; LINE Element Degenerate Primers; PCR Programmes.
One-Step Isolation of Plant DNA Suitable for PCR Amplification One step extraction for isolation of plant DNA. DNA suitable for amplification by PCR can be produced from leaf material smaller than 0.3 mm2 in less than 20 min & no tube changes. Method was tested on several plant species. Method was found to extract DNA that could be amplified without any further purification or treatment. The isolated DNA was amplified using a universal chloroplast primer set. The method was validated by comparing size of PCR products generated using standard DNA isolation.
Plant DNA Extraction Protocol A. thaliana has a very small haploid genome and this makes obtaining DNA somewhat difficult. The most notable problem is that DNA is usually contaminated with polysaccharide which inhibit restriction enzymes as well as other DNA modifying enzymes. This problem is most easily solved by using young plants which have not accumulated as much polysaccharide as older plants. The best results are obtained with plants that are two to three weeks post germinated.
Preparation of RNA from Plant Tissue Using Trizol Protocol is based on the standard Trizol protocol for the purification of RNA from animal cells using Trizol (Purification of RNA from Animal Cells using Trizol). In this version, adapted for use with plant tissues, a high-salt isopropanol precipitation step has been added to precipitate RNA selectively, while maintaining polysaccharides and proteoglycans in solution.
Whole-Mount DAPI Staining and Measurement of DNA Content in Plant Cells During development many plant cells undergo endoreduplication, whereby ploidy increases to a multiple of the normal 2C content. For eg., trichome development is accompanied by an increase in ploidy to 32C, indicating that trichome cells undergo four rounds of endoreduplication. Protocol describes DNA levels, and hence developmental progress in the corresponding cells, are measured by staining the DNA with a fluorescent marker and then quantifying the fluorescence of individual nuclei.
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