Abstract
Medicinal mushroom is defined as mushrooms used in medicine or medical research. Isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including Polymerase Chain Reaction (PCR), endonuclease restriction digestion, Southern blot analysis, and genomic library construction. The most important and prerequisite towards reliable molecular biology work is the total genomic DNA of a sample must be in good quality. Five freshly samples of medicinal mushroom were used in this work known as Auriculariapolytricha, Lentinus edode, Pleurotus sayorcaju, Sczhizopyllum commune and Ganodermalucidum. 5 mg of each sample were used to extraction the DNA, prepared in 3 replications and repeated twice. PCR based technique by using ISSR markers were used in checking the amplification ability of the total genomic extraction. A standard Doyle and Doyle protocol for genomic DNA extraction was modified in optimizing the total genomic DNA from the medicinal mushroom.The modification parameters were percentage of CTAB, incubation period and temperature. The results reveal that each sample required a certain combinations of time and period of incubation. Besides, percentage of CTAB in the buffer was found significant in giving a high yielding of extracted total genomic DNA. The extracted total genomic DNA from the medicinal mushroom yielded
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Mohamad, Azhar;
Mohd Mustafa, Muhammad Hussaini;
Azhari Noor, Muhammad Hanif;
Rashid, Rosnani Abdul;
Hasan Mutaat, Hasan Hamdani;
Meskom, Meswan;
Awang, Mat Rasol
[1]
- Malaysian Nuclear Agency, Bangi, Kajang, Selangor (Malaysia), Agroscience and Biotechnology Div.
Citation Formats
Mohamad, Azhar, Mohd Mustafa, Muhammad Hussaini, Azhari Noor, Muhammad Hanif, Rashid, Rosnani Abdul, Hasan Mutaat, Hasan Hamdani, Meskom, Meswan, and Awang, Mat Rasol.
A CTAB Procedure Of Total Genomic DNA Extraction For Medicinal Mushrooms.
Malaysia: N. p.,
2014.
Web.
Mohamad, Azhar, Mohd Mustafa, Muhammad Hussaini, Azhari Noor, Muhammad Hanif, Rashid, Rosnani Abdul, Hasan Mutaat, Hasan Hamdani, Meskom, Meswan, & Awang, Mat Rasol.
A CTAB Procedure Of Total Genomic DNA Extraction For Medicinal Mushrooms.
Malaysia.
Mohamad, Azhar, Mohd Mustafa, Muhammad Hussaini, Azhari Noor, Muhammad Hanif, Rashid, Rosnani Abdul, Hasan Mutaat, Hasan Hamdani, Meskom, Meswan, and Awang, Mat Rasol.
2014.
"A CTAB Procedure Of Total Genomic DNA Extraction For Medicinal Mushrooms."
Malaysia.
@misc{etde_22410678,
title = {A CTAB Procedure Of Total Genomic DNA Extraction For Medicinal Mushrooms}
author = {Mohamad, Azhar, Mohd Mustafa, Muhammad Hussaini, Azhari Noor, Muhammad Hanif, Rashid, Rosnani Abdul, Hasan Mutaat, Hasan Hamdani, Meskom, Meswan, and Awang, Mat Rasol}
abstractNote = {Medicinal mushroom is defined as mushrooms used in medicine or medical research. Isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including Polymerase Chain Reaction (PCR), endonuclease restriction digestion, Southern blot analysis, and genomic library construction. The most important and prerequisite towards reliable molecular biology work is the total genomic DNA of a sample must be in good quality. Five freshly samples of medicinal mushroom were used in this work known as Auriculariapolytricha, Lentinus edode, Pleurotus sayorcaju, Sczhizopyllum commune and Ganodermalucidum. 5 mg of each sample were used to extraction the DNA, prepared in 3 replications and repeated twice. PCR based technique by using ISSR markers were used in checking the amplification ability of the total genomic extraction. A standard Doyle and Doyle protocol for genomic DNA extraction was modified in optimizing the total genomic DNA from the medicinal mushroom.The modification parameters were percentage of CTAB, incubation period and temperature. The results reveal that each sample required a certain combinations of time and period of incubation. Besides, percentage of CTAB in the buffer was found significant in giving a high yielding of extracted total genomic DNA. The extracted total genomic DNA from the medicinal mushroom yielded from 39.7 ng/ μl to 919.1 ng/ μl. The different yield among the samples found to be corresponded to polysaccharide content in the medicinal mushrooms. The objective of this works is to optimize total genomic DNA extraction of medicinal mushrooms towards a high quality intact genomic DNA for molecular activities. (author)}
place = {Malaysia}
year = {2014}
month = {Jul}
}
title = {A CTAB Procedure Of Total Genomic DNA Extraction For Medicinal Mushrooms}
author = {Mohamad, Azhar, Mohd Mustafa, Muhammad Hussaini, Azhari Noor, Muhammad Hanif, Rashid, Rosnani Abdul, Hasan Mutaat, Hasan Hamdani, Meskom, Meswan, and Awang, Mat Rasol}
abstractNote = {Medicinal mushroom is defined as mushrooms used in medicine or medical research. Isolation of intact, high-molecular-mass genomic DNA is essential for many molecular biology applications including Polymerase Chain Reaction (PCR), endonuclease restriction digestion, Southern blot analysis, and genomic library construction. The most important and prerequisite towards reliable molecular biology work is the total genomic DNA of a sample must be in good quality. Five freshly samples of medicinal mushroom were used in this work known as Auriculariapolytricha, Lentinus edode, Pleurotus sayorcaju, Sczhizopyllum commune and Ganodermalucidum. 5 mg of each sample were used to extraction the DNA, prepared in 3 replications and repeated twice. PCR based technique by using ISSR markers were used in checking the amplification ability of the total genomic extraction. A standard Doyle and Doyle protocol for genomic DNA extraction was modified in optimizing the total genomic DNA from the medicinal mushroom.The modification parameters were percentage of CTAB, incubation period and temperature. The results reveal that each sample required a certain combinations of time and period of incubation. Besides, percentage of CTAB in the buffer was found significant in giving a high yielding of extracted total genomic DNA. The extracted total genomic DNA from the medicinal mushroom yielded from 39.7 ng/ μl to 919.1 ng/ μl. The different yield among the samples found to be corresponded to polysaccharide content in the medicinal mushrooms. The objective of this works is to optimize total genomic DNA extraction of medicinal mushrooms towards a high quality intact genomic DNA for molecular activities. (author)}
place = {Malaysia}
year = {2014}
month = {Jul}
}