Rapid and reliable extraction of genomic DNA from various wild-type and transgenic plants

BMC Biotechnology
Tae-Jin Kang, Moon-Sik Yang

Abstract

DNA extraction methods for PCR-quality DNA from calluses and plants are not time efficient, since they require that the tissues be ground in liquid nitrogen, followed by precipitation of the DNA pellet in ethanol, washing and drying the pellet, etc. The need for a rapid and simple procedure is urgent, especially when hundreds of samples need to be analyzed. Here, we describe a simple and efficient method of isolating high-quality genomic DNA for PCR amplification and enzyme digestion from calluses, various wild-type and transgenic plants. We developed new rapid and reliable genomic DNA extraction method. With our developed method, plant genomic DNA extraction could be performed within 30 min. The method was as follows. Plant tissue was homogenized with salt DNA extraction buffer using hand-operated homogenizer and extracted by phenol:chloroform:isoamyl alcohol (25:24:1). After centrifugation, the supernatant was directly used for DNA template for PCR, resulting in successful amplification for RAPD from various sources of plants and specific foreign genes from transgenic plants. After precipitating the supernatant, the DNA was completely digested by restriction enzymes. This DNA extraction procedure promises simplicity, speed, a...Continue Reading

References

Jul 11, 1995·Nucleic Acids Research·J J SteinerL F Elliott
Feb 12, 1998·Nucleic Acids Research·S M Aljanabi, I Martinez
Apr 12, 2001·Plant Physiology·K A LutzP Maliga
Mar 2, 2006·Journal of Agricultural and Food Chemistry·Ranjana SharmaTim A McAllister
Mar 1, 1985·Plant Molecular Biology·S O Rogers, A J Bendich
Dec 18, 2013·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Donghua GuoDongbo Sun

❮ Previous
Next ❯

Citations

May 13, 2009·Journal of Applied Genetics·I AhmedB Mirza
Jul 12, 2008·Molecular Biotechnology·Maria OszvaldMoon-Sik Yang
Sep 12, 2007·Advances in Neonatal Care : Official Journal of the National Association of Neonatal Nurses·UNKNOWN American Academy of PediatricsUNKNOWN Fetus and Newborn Committee
Apr 4, 2012·Advances in Neonatal Care : Official Journal of the National Association of Neonatal Nurses·Kimberly A Allen
Feb 27, 2013·Pediatric Critical Care Medicine : a Journal of the Society of Critical Care Medicine and the World Federation of Pediatric Intensive and Critical Care Societies·Xavier DurrmeyerRicardo Carbajal
Jan 22, 2010·Pediatric Research·Xavier DurrmeyerPeter C Rimensberger
Jun 15, 2007·Journal of Biochemistry and Molecular Biology·Young Geun YangDoo-Sik Kim
Feb 8, 2007·Biotechnology Journal·Astha VarmaNeeta Shrivastava
Oct 1, 2013·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Chuandeng YiMinghong Gu
Jul 11, 2013·World Journal of Microbiology & Biotechnology·N WatanarojanapornN Teaumroong
Mar 8, 2011·Archives of Pharmacal Research·Young Hun ShimHyoung Jin Kim
Oct 15, 2013·Biotechnology Progress·Edgar García LópezHerminia Guadalupe Martínez Rodríguez
Feb 5, 2015·The Plant Journal : for Cell and Molecular Biology·Reena NarsaiBrian J Atwell

❮ Previous
Next ❯

Methods Mentioned

BETA
PCR
transgenic
electrophoresis

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.