) Gel filtration chromatography on Sephadex G50 b.SDS-PAGE
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With the aid of diagrams, explain the processes and principles of each of the following
methods:
(a) Gel filtration chromatography on Sephadex G50
b.SDS-PAGE
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- What concentration of ethanol is considered optimum for disinfectant? Why not 100%?Please explain why is 70% ethanol used as an effective disinfecting solution in labs? Why not 95% ethanol? Please include references.How would you make two-fold serial dilutions such that the last tube is a 1:32 dilution of the original, concentrated material? Assume that you need to have at least 500 µl of each dilution, and you should be able to perform the dilutions in microfuge tubes with a maximum capacity of 1.5 ml.
- Explain how to prepare 250 ml liquid and 500 ml solid form separately from LB (Luria Bertani) medium whose components and concentrations are given below. Yeast extract 5 g/L Peptone 10 g/L NaCl 10 g/L Agar 1.5 g/Ldescribe each dosage form and enumerate and describe each kind. EMULSIONS-3 KINDS SOLUTIONS-3 KINDSWhen is sterilization by filtration preferred over sterilization by heat? Why? What makes heat an effective sterilizing agent? Why are the conditions required for sterilization greater for dry heat (160-180°C for 1-2hrs) than for steam (121°C for 15min)?
- In a mixture of five proteins listed, draw an elution profile (Absorbance vs. mL eluted, arbitrary) for the purification of the listed proteins on a gel filtration chromatography resin: cytochrome c (pI = 5.4; Mr = 13,000), immunoglobulin G (pI = 7.3; Mr = 145,000), ribonuclease A (pI = 9.6; Mr = 13,700), RNA polymerase (pI = 6.3; Mr = 450,000), human serum albumin (pI = 5.4; Mr = 68,500). Label your elution peaks. Draw a sketch of an SDS-PAGE, reflecting the mobility of the above mixture as they elute from the column. Label you protein bands.Which stationary phase is better in separation of the components of moringa extract for column chromatography? Sodium Bicarbonate or Silica Gel? And Explain.how does the diameter of the column affect the flow rate of the elution in gel filtration chromatography?
- Why Purified synthetic alcohol is preferred for level vial?In the traditional alkaline lysis method of Solution 1, what is the purpose of Glucosel?The plate with 45 colonies was inocculated with 100 µL Volume of a 10^-8 Dilution. Given the formula OCD = CFU/V x D, what as the Original Colony Density?