Preview

Isolation of Rna

Good Essays
Open Document
Open Document
298 Words
Grammar
Grammar
Plagiarism
Plagiarism
Writing
Writing
Score
Score
Isolation of Rna
Isolation of yeast RNA

Methods

The experiment was first started through mixing of 3.0 g dry yeast, 5.0 ml of 1% NaOH and 25.0 ml distilled water in 100 ml beaker. The resulting mixture was heated in a boiling water bath for 15 minutes while being stirred. The suspension was strained using cheesecloth and the obtained filtrate was obtained and collected in the beaker. This was then centrifuged at maximum speed for about 10-15 minutes. Centrifugation, which uses the idea of gravity, break up the cells and leads to the sedimentation of the largest particles present, the nuclei. Increasing the centrifugal speed brings about the sedimentation of first the cytoplasmic large and small granules. RNA was obtained from the cytoplasmic fractions.

After centrifugation, the supernatant was collected while the residue was discarded. Glacial acetic acid was added in the supernatant until it became slightly acidic. The obtained solution was again centrifuged and filtered through the cheesecloth. Centrifugation and filtration was performed many time until the supernatant became clear.
The supernatant, was placed in a boiling water bath on which it is evaporated to approximately 5 ml. The 5 ml mixture was cooled to 40oC and 10 ml acidified 95% ethanol was included with vigorous stirring.

The resulting mixture was placed in an ice bath for at least thirty minutes. This was again centrifuged for 5 minutes at 6000 rpm and the supernatant was discarded. The formed residue was washed with 1ml of 95% ethanol and twice with a small amount of water. The washed crude RNA was transferred in an evaporating dish and was quantitatively determined for its weight. This stock RNA was then added with 0.14 M Tris-HCl buffer to form 10 ml of 10%(w/v) stock RNA

You May Also Find These Documents Helpful

  • Good Essays

    CHEM 2070 LAB

    • 604 Words
    • 3 Pages

    After trying to decant the solution, and attempting to keep the precipitate in the beaker, which was difficult to do as some of the precipitate left the solution along with the hot supernatant liquid in the heated solution, 20 mL of distilled water was added to the precipitate.…

    • 604 Words
    • 3 Pages
    Good Essays
  • Satisfactory Essays

    In this experiment, you will model the effects of mutations on the genetic code. Some mutations cause no structural or functional change to proteins while others can have devastating affects on an organism.…

    • 624 Words
    • 3 Pages
    Satisfactory Essays
  • Powerful Essays

    The aim of the experiment was to test for the presence of DNA, RNA, protein and glycogen in the cytoplasm and the nucleus of bovine liver cells. From the findings of the results the distribution of these macromolecules can be shown within the liver cell. This was carried out by undertaking qualitative experiments, where the observation of a colour change was noted and a quantitative experiment, where numerical data was recorded from the measurements of light absorbance.…

    • 3513 Words
    • 15 Pages
    Powerful Essays
  • Good Essays

    Competing Nucleophiles Lab

    • 2607 Words
    • 11 Pages

    40 g of ice and approximately 30 ml of sulfuric acid is cautiously added to a 100 mL beaker respectively. Weigh 7.6 g of ammonium chloride and 14.0 g of ammonium bromide and place it in another beaker, crushing the lumps until a powdery mixture remains. The powdery mixture is then transferred to a 125 mL Erlenmeyer flask. Add the ammonium salts into the sulfuric acid mixture. Heat is applied to dissolve the salt. Once the solution has fully dissolved, allow it to cool until the warm substance is touchable. A prolonged waiting period can cause the solution to recrystallize and this process would have to be repeated again. Place 6.0 mL of water in a 15 mL centrifuge tube marking the tube at the 6.0 mL level and replace the water with the solvent-nucleophile mixture. The tube is set aside for the t-butyl experiment. The remaining mixture is placed in a 25 mL round bottom flask.…

    • 2607 Words
    • 11 Pages
    Good Essays
  • Good Essays

    benzoate was added, the cold nitric acid/sulfuric acid mixture was added to the conical vial…

    • 768 Words
    • 3 Pages
    Good Essays
  • Satisfactory Essays

    Chem Lab

    • 757 Words
    • 4 Pages

    From conical vial contents were then transferred to a clean centrifuge tube. The mixture was allowed to separate into two layers. Newly formed aqueous layer was removed with a plastic pipette and stored in a 50 ml beaker.…

    • 757 Words
    • 4 Pages
    Satisfactory Essays
  • Good Essays

    6.03 Calorimetry Lab

    • 446 Words
    • 2 Pages

    The calcium dissolved really quickly and there were bubbles and foams formed. We then heated the beaker with a hot plate for 2 to 3 minutes, equally distributed the supernatant solution into two test tubes, but avoid pouring the solid. After centrifuging the samples, we added extra HCl solution into the test tubes, there wasn’t any bubbling occurs, so we could continue and put the mixtures into the centrifuge again. And then put the NaCO3 solution into the beaker and wait to allow the CaCO3 precipitate to form. We then tested whether the supernatant solution is still acidic with a piece of red litmus paper, filter the liquid and we had some CaCO3. Dried the filter paper and CaCO3 in the oven, then we broke the CaCO3 into small particles with a glass rod, and dried it one more time. Let it cool and repeat heating, cooling and weighing it until the masses differ between two results are less than…

    • 446 Words
    • 2 Pages
    Good Essays
  • Good Essays

    The purpose of this lab was to evaluate our skills of decanting a supernatant liquid without losing the solid and successful completion of a series of reactions. This was done through five chemical reactions involving copper. In this lab, elemental copper was put through five different chemical reactions in order to convert it into different compounds. By the end of the fifth reaction, the copper was back to its elemental state.…

    • 842 Words
    • 4 Pages
    Good Essays
  • Satisfactory Essays

    Rna Primase

    • 206 Words
    • 1 Page

    DNA primase is an enzyme involved in the copping of DNA and is a type of RNA polymerase. Primase speed up the synthesis of a short RNA (or DNA in some organisms [1]) a part called a primer complementary to a ssDNA template. Primase is of key importance in DNA replication because no known replicative DNA polymerases can begin the synthesis of a DNA strand without an initial RNA or DNA primer. After this elongation, the RNA piece is removed by a 5' to 3' exonuclease and refilled with DNA. primase uses a phosphotransfer domain for the transfer coordination of metals, which makes it distinct from other polymerases. The side subunits include a NH2 and COOH terminal made of alpha helixes and beta sheets. The NH2 terminal reacts with a zinc binding…

    • 206 Words
    • 1 Page
    Satisfactory Essays
  • Satisfactory Essays

    DNA and RNA

    • 352 Words
    • 1 Page

    Deoxyribonucleic acid and ribonucleic acid also know as DNA and RNA a closely related molecules that participate in transmitting and expressing genetic information. Both the DNA and the RNA have molecular chains containing alternating units of sugar and phosphate. Now each sugar unit has nitrogen containing molecules called nucleotide bases that hand off of them. The different sugar units in DNA and RNA are responsible for the differences between the two biochemicals. To contrast DNA and RNA you need to look at the physical structure of the two. DNA and RNA are identical except that on carbon binds to a hydrogen atom instead of a hydroxyl group. This difference means that two strands of DNA can form a double-helix structure while the RNA remains as a single strand. With that being said DNA 's double-helix structure is very stable, giving it the ability to encode information for a long time. Moreover, the cell creates RNA as needed during the process of transcription, but DNA is self-replicating.…

    • 352 Words
    • 1 Page
    Satisfactory Essays
  • Good Essays

    1. OK, so remember that back in the day, we had protein and nucleic acid resulting from a centrifugation…Well, now we're dealing with the protein portion, which is solid…

    • 3884 Words
    • 16 Pages
    Good Essays
  • Powerful Essays

    Lidocaine Synthesis

    • 1214 Words
    • 4 Pages

    An ice water bath for 100 mL of water was prepared. It was noted that gloves had to be put on at this time. 3.0 mL of 2,6-dimethylaniline in a 10 mL graduated cylinder. Then, a larger cylinder was used to measure out 15 mL of glacial (concentrated) acetic acid. This was poured into a 125 mL Erlenmeyer flask. Both graduated cylinders were rinsed thoroughly with acetone several times. The 2,6-dimethylaniline was added via a Pasteur pipet to the acetic acid. The graduated cylinder was rinsed with acetone and left to air dry. 2 mL of 2-chloroacetyl chloride was measured and poured into the Erlenmeyer flask. A thermometer was placed to record any temperature change during this process. A large graduated cylinder was used to measure out 25 mL of .333 M Sodium Acetate directly to the Erlenmeyer flask. The temperature increased from 30 to 32 degrees Celsius. 60 mL of…

    • 1214 Words
    • 4 Pages
    Powerful Essays
  • Powerful Essays

    Rna World

    • 3500 Words
    • 14 Pages

    Editors: John F. Atkins, Raymond F. Gesteland, and Thomas R. Cech Additional Perspectives on RNA Worlds available at www.cshperspectives.org Copyright # 2012 Cold Spring Harbor Laboratory Press; all rights reserved; doi: 10.1101/cshperspect.a006742 Cite as Cold Spring Harb Perspect Biol 2012;4:a006742…

    • 3500 Words
    • 14 Pages
    Powerful Essays
  • Good Essays

    Memantine-Hcl Lab Report

    • 1846 Words
    • 8 Pages

    Some deionized water was added and drug powder was dissolved in water. Then required amount of DCM was added. The pH adjustment was done by adding a few drops of TEA, afterwards two distinctive phases were formed. pH value of supernatant was measured quickly (pH-meter showed number 11-12). The reaction flask was heated and shaken for 30 minutes, followed by decantation to obtain base Memantine (without Hcl). The reaction flask was connected to rotary evaporator equipped with vacuum pump to remove excess solvent (i.e.,…

    • 1846 Words
    • 8 Pages
    Good Essays
  • Better Essays

    5. Biological Centrifugation (Spin biological), by D. Rickwood, J.M. Graham (2001). [Online]. Available at: http://www.coleparmer.com/TechLibraryArticle/910…

    • 1541 Words
    • 7 Pages
    Better Essays