Molecular Biology Paper

Lab Report 1 Introduction A cells plasma tissue layer is cognize to be selectively pervious. This implies that the tissue layer is selective on what substances can follow out in and out of the cell. There be 2 methods of transport that encounter done the plasma tissue layer. One method of transport is c every(prenominal)ed active make for which uses ATP energy to transport substances done the tissue layer. The other method is called unresisting cover which does non require the use of ATP energy. During pull finishedive processes, molecules are transported through the membrane by differences in tautness or tweet between the inside and outside of the cell.Two important types of passive process are sp take aiming and filtration. Every cell in the human corpse uses diffusion as an important transport process through its selectively permeable membrane. During diffusion, molecules that are small enough to pass through a membranes revolve abouts or molecules that can dissolve in the lipoid section of a membrane move from an part of higher in decennarytness to an area of dismay concentration. The kinetic energy that all molecules possess is the motive force in diffusion. Facilitated diffusion occurs when molecules are too deep to pass through a membrane or are lipid water-insoluble.In this process, bearer protein molecules located in the membrane combine with solutes and transport them tidy sum the concentration gradient. Filtration is another type of passive process and, unlike diffusion this is not a selective process. The insistency gradient on individually side of the membrane as well as the membrane pore size depends on the nume ordinate of solutes and fluids in the trickle. During filtration, water and solute molecules pass through a membrane from an area of higher hydrostatic pressure to an area of lower hydrostatic pressure.This means that water and solutes would pass through a selectively permeable membrane along the pressu re gradient. To gain a better understanding of a cells selectively permeable membrane and the passive processes of simple diffusion, facilitated diffusion, and filtration, three look intos were conducted. Materials and Methods Activity 1 Simulating Dialysis (Simple Diffusion) Materials ? cardinal glass beakers ? quad dialysis membranes 20 (MWCO), 50 (MWCO), coulomb (MWCO), and cc (MWCO) ? membrane pallbearer ? membrane barrier ? four solutes NaCl, carbamide, Albumin, and Glucose beginning dispenser ? deionized water ? timepiece ? beaker flush This experiment was conducted first by placing the 20 (MWCO) dialysis membrane into the membrane toter. The membrane holder joined the deuce glass beakers one on the left side and one on the right side. Then, 9. 00 mM of NaCl concentration was dole out into the left beaker. Deionized water was dispensed in the right beaker. When the horologe was started, the barrier that surrounded the membrane holder was lowered to impart the table of contents of sepa come outly beaker to come in contact with the membrane.After the 60 proceedings of compressed time elapsed, results were read and recorded. Finally, each beaker was past flushed for readiness of the coterminous experiment run. These exact steps were followed using each dialysis membrane size (20, 50, 100, and two hundred) as well as with each solute (NaCl, urea, Albumin, and Glucose). There were a total of sixteen runs in this experiment. Activity 2 Simulating Facilitated Diffusion Materials ? two glass beakers ? membrane builder ? membrane holder ? glucose concentration ? solution dispenser ? deionized water ? timer beaker flush In this experiment, the first step was to do the glucose carrier to ergocalciferol in order to correctly build the membrane. Next, a membrane was built in the membrane builder by inserting 500 glucose carrier proteins into it. Then, the newly built membrane was placed into the membrane holder that joined the two glass beakers. Th e two glass beakers were joined on the left and right sides of the membrane holder. After that, 2. 00 mM of glucose concentration was dispensed into the left beaker. The right beaker was filled with deionized water.The barrier around the membrane holder dropped when the timer was started. After 60 legal proceeding of compressed time elapsed, the results were read and recorded. Finally, both glass beakers were flushed to prepare for the next experimental runs. The higher up mentioned steps were repeated by increasing the glucose concentration to 8. 00. Both the 2. 00 mM and the 8. 00 mM glucose concentration solution were tested using membranes built with 500, 700, and 900 glucose carrier proteins. There were a total of six experimental runs. Activity 4 Simulating Filtration Materials ? two glass beakers membrane holder ? 4 dialysis membranes 20 (MWCO), 50 (MWCO), 100 (MWCO), and 200 (MWCO) ? 4 solutions Na+Cl? , Urea, glucose, and pulverized charcoal grey ? solution dispenser ? pressure unit ? timer ? filtration enumerate indicator ? membrane residue analysis analyzer ? beaker flush In the final experiment, the two glass beakers were placed one on top off of the other with the membrane holder between them. The pressure unit that rested on the top beaker was used for forcing the solution from the top beaker through the selected membrane and into the toilet beaker.The bottom beaker contained nothing however, the filtration rate indicator was attached to it from one side. The experiment began by placing the 20 (MWCO) dialysis membrane into the membrane holder. Then, 5. 00 mg/ml of each of the chase solutions Na+Cl? , Urea, glucose, and powdered charcoal were dispensed into the top beaker. The pressure unit was adjusted to 50 mmHg of pressure. The timer was set to 60 minutes of compressed time and when the timer started, the membrane holder retracted. The solution consequently flowed through the membrane and into the beaker underneath.When the timer stopp ed, the membrane was then placed in the membrane residue analysis analyzer. The results were read and recorded and the beakers were flushed for the next experimental runs. All the above steps were repeated using the 50 (MWCO), 100 (MWCO), and 200 (MWCO) membranes. Results Table 1 Activity 1 Simulating Dialysis (Simple Diffusion) notice Solutes that were able to diffuse into the right beaker are indicated by a +. Solutes that were not able to diffuse into the right beaker are indicated by a -. tissue layer (MWCO) Solute (9. 0 mM) (Pore Size) NaCl Urea Albumin Glucose 20 50 + 100 + 200 + + graph 1 Activity 2 Simulating Facilitated Diffusion Glucose Transport regularise (mM/min) pic Table 2 and 3 Activity 4 Simulating Filtration Table 2 Solute Residue Presence in the Membrane Key If solute residue was symbolize on the membrane, it is indicated by a +. If solute residue was not present on the membrane, it is indicated by a .Membrane (MWCO) Solute 20 50 100 200 NaCl + + + + Urea + + + + Glucose + + + + powdered Charcoal + + + + Table 3 Filtration Rate and nub of Solute Detected in Filtrate Membrane (MWCO) Solute 20 50 100 200 Filtration Rate (ml/min) 1 2. 5 10 NaCl in tense (mg/ml) 0 4. 81 4. 81 4. 81 Urea in filtrate (mg/ml) 0 0 4. 74 4. 74 Glucose in filtrate (mg/ml) 0 0 0 4. 9 Powdered Charcoal (mg/ml) 0 0 0 0 Discussion The first lab experiment, Simulating Dialysis (Simple Diffusion), demonstrated how only if certain molecules pass through a selectively permeable membrane down its concentration gradient. The four membranes utilized in this experiment consisted of each one world different in pore size (MWCO). The smallest pore-sized membrane was 20 (MWCO), and the largest was 200 (MWCO). The solutes that were tested in this experiment were NaCl, Urea, Albumin, and Glucose.The first solute tested, NaCl, showed that with a 20 (MWCO) membrane, no diffusion occurred into the right beaker. (Table 1) Th e NaCl molecules were evidently too large to pass through the 20 (MWCO) membrane because its pores were too small. Membranes 50, 100, and 200 (MWCO) did allow the NaCl to pass through. (Table 1) One of the reasons this occurred is because the pores in the above mentioned membranes were large enough to rent the passage of the NaCl molecules. The other reason diffusion occurred is because the NaCl molecules moved down its concentration gradient and into the beaker filled with deionized water. For all three membranes, equilibrium was reached in ten minutes at an average out diffusion rate of 0. 0150 mM/min.As for the solute Urea, the experiment conducted showed that no diffusion occurred with all four membranes. (Table 1) Urea should give up passed through membranes 100 (MWCO) and 200 (MWCO) for the reasons that its molecules are small enough and Urea is also soluble. This experiment showed that none of the Albumin molecules diffused through all of the four membranes tested. (Table 1) This is because the Albumin molecules were too large to pass through the pores of all four membranes. The final solute tested in this experiment, Glucose, showed that the molecules only diffused through the 200 (MWCO) membrane. (Table 1) Equilibrium was reached in thirty-seven minutes at an average diffusion rate of 0. 0040 mM/min.The Glucose molecules were too large to diffuse through the 20 (MWCO), 50 (MWCO), and 100 (MWCO) membranes. The second experiment, Simulating Facilitated Diffusion, explained how carrier protein molecules in the membrane effectively transported molecules that are too large or are insoluble to diffuse through the membrane. The carrier proteins in this experiment were glucose carriers and the solution was a 2. 00 (mM) and an 8. 00 (mM) glucose concentration. The 2. 00 (mM) glucose concentration was tested first with the 500 glucose carrier protein membrane then the 700 and 900 glucose carrier protein membranes. The glucose transport rate for the membrane with 500 glucose carrier proteins was 0. 0008 (mM/min). Graph 1) The membrane with 700 glucose carrier proteins showed a rate of 0. 0010 (mM/min) and the 900 glucose carrier proteins membrane had a rate of 0. 0012 (mM/min). (Graph 1) The 8. 00 (mM) glucose concentration also showed and increase in glucose transport rate with membranes that contained more glucose carrier proteins. The membrane with 500 glucose carrier proteins showed a rate of 0. 0023 (mM/min). (Graph 1) Membranes that had 700 and 900 glucose carrier proteins showed a rate of 0. 0031 and 0. 0038 (mM/min). (Graph 1) These results show that with an increase in amount of glucose carrier proteins in the membranes, transport of the glucose molecules in the concentration is more effective.A higher concentration of glucose (8. 00 mM) also increases the rate of glucose transport in a membrane with the aforementioned(prenominal) amount of glucose carrier proteins as a lower glucose concentration (2. 00). The final experimen t, Simulating Filtration, four different solutes were forced through four membranes that contained separate pore sizes by the use of hydrostatic pressure. After each experimental run was conducted, the membrane analyses showed that residue from all four solutes were detect on each membrane. (Table 2) This indicates that some solutes did not filter through the membrane. The filtration rate (ml/min) increased as membranes with bigger pores were utilized.This happened because the solute molecules were able to transport through a particular membrane at a faster rate being that the membranes pores were larger. The filtrate in the bottom beaker was analyzed and no solutes were detected with the 20 (MWCO) membrane. (Table 3) With the 50 (MWCO) membrane, only NaCl was detected in the filtrate at 4. 81 (mg/ml). (Table 3) The 100 (MWCO) membrane showed to have NaCl at 4. 81 (mg/ml) and Urea at 4. 74 (mg/ml) present in the filtrate. (Table 3) Glucose and powdered charcoal were not present. The expiry membrane with pore size 200 (MWCO), had the solutes NaCl at 4. 81 (mg/ml), Urea at 4. 74 (mg/ml), and Glucose at 4. 39 (mg/ml) detected in the filtrate. (Table 3) Powdered charcoal was not detected in this filtrate. Table 3) The molecules in powdered charcoal were too large to pass through any of the membranes tested. The 20 (MWCO) membrane pores were too small to allow any solute molecules to pass through. The membranes that contained larger pores allowed the solutes with larger pores pass through. The amounts (mg/ml) of the same solute detected in the filtrate were the same for each membrane. (Table 3) This is because the pressure that was released into the top beaker remained at 50 (mmHg) for all experiment runs. References Marieb, Elaine N. , Mitchell, Susan J. (2008). Exercise 5B. Human Anatomy & Physiology Laboratory Manual Ninth mutation (pp. PEx-5 PEx-13). San Francisco, California Pearson Benjamin Cummings.