Isolation of Genomic Dna from Animal Tissues
LAB REPORT 4
Isolation of genomic DNA from animal tissues
Introduction A rapid expansion of DNA analysis in medical, biotechnological and basic research has created the need for simple and efficient commercial methods for isolating genomic DNA. Many different methods and technologies are available for the isolation of genomic DNA. In general, all methods involve disruption and lysis of the starting material followed by the removal of proteins and other contaminants and finally recovery of the DNA. Removal of proteins is typically achieved by digestion with proteinase K, followed by salting-out, organic extraction, or binding of the DNA to a solid phase support (either anion-exchange or silica technology). DNA is usually recovered by precipitation using ethanol or isopropanol. The choice of a method depends on many factors.The required quantity and molecular weight of the DNA, the purity required for downstream applications, and the time and expense.The separation of DNA from cellular components can be divided into four stages. Disruption, Lysis, Removal of proteins and contaminants and Recovery of DNA.
Objective 1. Learn how to isolate genomic DNA from animal tissue 2. To observe the phases of animal tissue in micro centrifuge.
Materials 1. Fresh tissue of animal ( fish ) 2. Mortal and pestle, 3. Petri dish 4. Lysis buffer 5. 1.5mL centrifuge tube 6. Phenol : chloroform (1:1) 7. Proteinase K 8. 3M NaOAc (ph 6.0) 9. Absolute ethanol 10. 70% ethanol 11. TE buffer / distilled water
Procedure 1) Fresh fish was cut to 0.5cm 2) Fish tissue washed with 70% alcohol 3) Wetted the tissue with a few drop of 70% of alcohol ( in petri dish) 4) Minced the tissue using mortal and pestle 5) Transferred minced tissue into a sterile 1.5mL centrifuge tube 6) Added 500ml of lysis buffer 7) Added 50ml of proteinase k 8) Incubated suspension (20min, 65 °C) 9) Mixed the
Isolation of genomic DNA from animal tissues
Introduction A rapid expansion of DNA analysis in medical, biotechnological and basic research has created the need for simple and efficient commercial methods for isolating genomic DNA. Many different methods and technologies are available for the isolation of genomic DNA. In general, all methods involve disruption and lysis of the starting material followed by the removal of proteins and other contaminants and finally recovery of the DNA. Removal of proteins is typically achieved by digestion with proteinase K, followed by salting-out, organic extraction, or binding of the DNA to a solid phase support (either anion-exchange or silica technology). DNA is usually recovered by precipitation using ethanol or isopropanol. The choice of a method depends on many factors.The required quantity and molecular weight of the DNA, the purity required for downstream applications, and the time and expense.The separation of DNA from cellular components can be divided into four stages. Disruption, Lysis, Removal of proteins and contaminants and Recovery of DNA.
Objective 1. Learn how to isolate genomic DNA from animal tissue 2. To observe the phases of animal tissue in micro centrifuge.
Materials 1. Fresh tissue of animal ( fish ) 2. Mortal and pestle, 3. Petri dish 4. Lysis buffer 5. 1.5mL centrifuge tube 6. Phenol : chloroform (1:1) 7. Proteinase K 8. 3M NaOAc (ph 6.0) 9. Absolute ethanol 10. 70% ethanol 11. TE buffer / distilled water
Procedure 1) Fresh fish was cut to 0.5cm 2) Fish tissue washed with 70% alcohol 3) Wetted the tissue with a few drop of 70% of alcohol ( in petri dish) 4) Minced the tissue using mortal and pestle 5) Transferred minced tissue into a sterile 1.5mL centrifuge tube 6) Added 500ml of lysis buffer 7) Added 50ml of proteinase k 8) Incubated suspension (20min, 65 °C) 9) Mixed the
References: 1. PubMed 2. Ramirez-Solis R, Rivera-Prez J, Wallace JD, Wims M, Zheng H, Bradley A. Genomic DNA microextraction: a method to screen numerous samples. Anal Biochem. 1992