Report Issue Data Analysis

Report Issue Data Analysis  Photograph the destained gel on an illuminated box each.  Create a standard curve of the protein standards plotting log of molecular weight against distance moved from the wells.  Using a standard curve of the protein standards calculate the size of the GFP purified in the: o Column purification samples o In the cell pellet lysates of the lb/ara/amp. You are only going to size the band(s) that is darker in the lb/ara/ amp sample. 5 o Prepare samples of each of the samples (tube 1,2,3 and lysate if you have it). o Heat all the samples at 95 o C for 10mins Cell Pellets o You will have two pellets from lab 9: ara and ara/amp o Add buffer to each tube as below Components Volume Pellet NUPAGE LDS Sample buffer 200ul Total volume 200ul o Aliquot 20 ul of ara and ara/amp two separate tubes labeled ‘heat’ o Aliquot 20 ul of ara and ara/amp two separate tubes labeled ‘no-heat’ o Incubate the ‘heat’ samples for 10 minutes at 95 o C Sample Loading o Use the pipette equipped with a protein gel loading tip to underlay the samples into the gel wells (see figure below). o Slowly draw the sample up into the tip as it will take time to fill due to the narrow bore. o Lower the tip to the bottom of the sample well and slowly pipet sample into well without contaminating another well with the sample. Gel 1 – Samples for gel lanes o Protein ladder – 5ul o Tube #1 – 10ul 6 o Tube #2 – 10 ul o Tube #3 – 10 ul o Lysate – 10 ul or leave blank o Protein ladder 5ul o Tube #1 – 15 ul o Tube#2 – 15ul o Tube #3 – 15 ul o Lysate – 15ul or leave blank Gel 2 – samples lanes o Protein ladder – 5ul o amp – heat o amp- no heat o ara/amp- heat o ara/amp – no heat o protein ladder – 5ul o amp – heat o amp- no heat o ara/amp- heat o ara/amp – no heat the first photo is Gel 1 , the second photo in Gel 2

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