Methodology In a 100-mL beaker, 3.0 g dry yeast was weighed and 5.0 mL 1% NaOH and 25.

0 mL distilled water were added. The solution was heated in boiling water for 15 minutes with occasional stirring. The suspension was filtered through cheesecloth and the filtrate was collected in a beaker. The filtrate was transferred into a 15-mL corning tube and centrifuged at maximum speed for 5 minutes. The supernatant was collected into another beaker and the residue was discarded. Glacial acetic acid was added dropwise to the supernatant until the supernatant became slightly acidic to litmus. The supernatant was then transferred into a 15-mL corning tube then centrifuged. The supernatant was collected and filtered through cheesecloth. The centrifugation and filtration process were repeated until the supernatant was clear. The supernatant was collected and evaporated over a boiling water bath to approximately 5 mL. The supernatant was then allowed to cool to about 40or lower. Into the cooled extract, 10 mL acidified 95% ethanol was added with vigorous stirring. The acidified 95% ethanol was prepared by adding concentrated HCl into 95% ethanol dropwise. Afterwards, acidity is checked with litmus paper. The mixture was then placed in an ice bath for around 5 minutes. Afterwards, the mixture was centrifuged for 5 minutes at 6000 rpm. The supernatant was decanted. The residue was washed with 2 mL 95% ethanol, then with 5 mL ether afterwards. The washings were decanted after centrifugation for 2 minutes. The washed crude RNA was transferred into a preweighed evaporating dish. The evaporating dish with the crude RNA was then weighed and transferred into a 15-mL corning tube. The weight and percentage recovery of RNA was calculated and determined. 10 mL of 10% (w/v) stock RNA solution was prepared by addition of 0.14 M Tris-HCl buffer pH 8.0. The crude RNA extract was stored in the refrigerator for future experiments. Introduction Ribonucleic acids (RNA) are responsible for protein synthesis. RNA is similar in many ways to deoxyribonucleic acid (DNA) except that the sugar moiety in RNA is a ribose while in DNA, it is a deoxyribose. Other than that, the pyrimidine nucleotide base in RNA is uracil instead of thymine. Most RNAs are single-stranded; some produce double stranded hybrids with DNA strands. For a particular cell to be considered a good source of RNA, it should have a high cytoplasmic to nuclear mass ratio since RNA is commonly found in the cytoplasm. One potential source of RNA is the yeast Saccharomyces cerevisiae which has approximately 4% RNA by mass. RNA extraction methods and tehcniques may vary depending on the source and the type of RNA to be isolated and extracted. The common types of RNA include mRNA, rRNA, and tRNA. The general methods for the extraction of RNA include (1) heating with alkali, (2) acid extraction at pH 4-5, (3) addition of acidified alcohol and (4) successive washing with alcohol and ether. Reagents 1% NaOH alkali lysis of cell Heating thermal lysis

Filtration through cheesecloth breaking down of membranes Centrifugation separate mo yung components ng cells Glacial acetic acid lowers down pH Acidified 95% ethanol less soluble nucleic acids sa ethanol Ice bath masma-eenhance yung precipitation; hindi madedenature yung RNA Concentrated HCl tanggal yung excess OHEther tinatanggal niya yung proteins and lipids 0.14 M Tris-HCl buffer pH 8.0 Description of extracted RNA The RNA extracted appears to be brownish powder or granules that are stuck to each other due to the 95% ethanol and ether washing. Theoretically, extracted RNA from Saccharomyces cerevisiae should appear ____________. Errors Recommendations