Sandbox 323: Difference between revisions
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The molecular weight of 3DS8 is proposed to be about 28 kDa. | The molecular weight of 3DS8 is proposed to be about 28 kDa, based on the length of the amino acid sequence. | ||
In the laboratory, the gene was transformed into ''E. coli'' bacteria, expressed and isolated by cell lysis and nickel column. The cell lysis buffer used was a sodium phosphate buffer. From the column, 10 elution fractions were collected. A Bradford assay with standards of 0.125, 0.25, 0.5, 0.75, and 1 mg/mL of Bovine Serum Albumin solution. The standards were measured on a UV-Vis spectrophotometer at 595 nm. Initially, a few of the elution fractions were out of the standard range of absorbance, so the samples were remade by diluting further. After the second round of absorbances was obtained, only elution 4 was still out of range. Then Beer's law (A=εbc) was used to determine the concentrations (mg/mL) of each elution fraction: E1=0.4026, E2=5.968, E3=5.434, E4=-0.1407, E5=0.5610, E6=0.4229, E7=0.03175, E8=0.1461, E9=0.2693, E10=0.2244. | In the laboratory, the gene was transformed into ''E. coli'' bacteria, expressed and isolated by cell lysis and nickel column. The cell lysis buffer used was a sodium phosphate buffer. From the column, 10 elution fractions were collected. A Bradford assay with standards of 0.125, 0.25, 0.5, 0.75, and 1 mg/mL of Bovine Serum Albumin solution. The standards were measured on a UV-Vis spectrophotometer at 595 nm. Initially, a few of the elution fractions were out of the standard range of absorbance, so the samples were remade by diluting further. After the second round of absorbances was obtained, only elution 4 was still out of range. Then Beer's law (A=εbc) was used to determine the concentrations (mg/mL) of each elution fraction: E1=0.4026, E2=5.968, E3=5.434, E4=-0.1407, E5=0.5610, E6=0.4229, E7=0.03175, E8=0.1461, E9=0.2693, E10=0.2244. | ||
