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• 20.109(F09): Mod 1 Day 8 FACS analysis
  ... sample of this cell type from a complex mixture by using a FACS machine. In addition to purification, the FACS machine can count the number of cells th ...rged droplets, each containing a single cell. The stream of droplets pass in front of a laser beam, and the scattered light is analyzed by a series of f
  5 KB (804 words) - 19:38, 28 July 2015
• 20.109(F09):DNA engineering "Progress Report"
  ...ualifying exams, but now they would like to hear what progress you've made in the lab over the last few months. The following guidelines have been adapte This assignment is due by 11:00 a.m. on the day you have lab. Please turn in your progress reports electronically by uploading them to the [http://stell
  7 KB (1,083 words) - 19:38, 28 July 2015
• 20.109(F09):DNA engineering "Memo"
  ...in which you argue either FOR or AGAINST taking the delta5/delta3 research in your division forward. This assignment is due by 11:00 a.m. on the day you have lab. Please turn in your memos electronically by uploading them to the [http://stellar.mit.edu/
  5 KB (747 words) - 19:38, 28 July 2015
• 20.109(F09):Lab tour
  ... these stations - all are fair game, as is the lab safety material covered in today's pre-lab lecture.'''</font color> ...ence ones. If time permits, you will read the absorbance of your dilutions in the spectrophotometer so transfer the volumes to cuvettes and do not throw
  27 KB (4,508 words) - 19:38, 28 July 2015
• 20.109(F09): Mod 2 Day 6 Readouts of DNA, Protein
  ...ression of the light sensing protein, Cph8, and the mutant versions of it. In general, detection depends on which antibody you choose, and the quality of ...ize one epitope, while other antibodies, called polyclonal antibodies, are in fact antibody pools that recognize multiple epitopes. We will be using a mo
  13 KB (2,115 words) - 19:38, 28 July 2015
• 20.109(F09): Mod 2 Day 2 Measuring system performance
  ...e output...then the cells might swim to the light rather than turn a color in response. ...nderstanding of the system we're working with is obviously needed. To look in the black box that senses light, we'll have to learn more about the two com
  11 KB (1,859 words) - 19:39, 28 July 2015
• 20.109(F10): Mod 2 Day 7 Readout of Protein
  ...ression of the light sensing protein, Cph8, and the mutant versions of it. In general, detection depends on which antibody you choose, and the quality of ...ize one epitope, while other antibodies, called polyclonal antibodies, are in fact antibody pools that recognize multiple epitopes.
  10 KB (1,613 words) - 21:00, 28 July 2015
• 20.109(F10):DNA engineering "Progress Report"
  ...ualifying exams, but now they would like to hear what progress you've made in the lab over the last few months. The following guidelines have been adapte This assignment is due by 11:00 a.m. Please turn in your progress reports electronically by uploading them to the [http://stell
  7 KB (1,131 words) - 21:00, 28 July 2015
• 20.109(F10):Lab tour
  ... these stations - all are fair game, as is the lab safety material covered in today's pre-lab lecture.'''</font color> ...ence ones. If time permits, you will read the absorbance of your dilutions in the spectrophotometer so transfer the volumes to cuvettes and do not throw
  27 KB (4,538 words) - 21:00, 28 July 2015
• 20.109(S11):Amplify aptamer-encoding DNA (Day1)
  ... screening, and has a heme binding affinity of 220 nM. Both were described in [http://www.ncbi.nlm.nih.gov/pubmed/17168539 this] 2006 paper. ...rage capabilities. We will make use of these capabilities more extensively in Module 2.
  10 KB (1,688 words) - 21:05, 28 July 2015
• 20.109(S11):Prepare DNA for cloning (Day3)
  ...etting it slowly cool - very similar to how we denatured RNA and cooled it in Module 1, except this time we want two strands to anneal rather than to for ... digests to understand how they work and why we had to do this one for you in the interests of time.
  17 KB (2,967 words) - 21:05, 28 July 2015
• 20.109(F11)
  ...ng, notebook keeping and publicly presenting your data. All of us involved in teaching 20.109 hope you will find it a satisfying challenge and an excitin *Agi's Monday office hour is cancelled. I'm in no shape to be breathing near anyone; I'm happy to discuss research proposa
  4 KB (629 words) - 21:07, 28 July 2015
• 20.109(F11):DNA engineering powerpoint summary
  *This assignment is due by 1PM. *This assignment can be '''no more than 10 slides total.'''
  3 KB (528 words) - 21:08, 28 July 2015
• 20.109(F11): Mod 1 Day 6 Lipofection & lab practical
  ...ld be transcribed by RNA polymerase. The polyadenylation sequence assists in the export and stability of the mRNA so that it gets translated by the ribo ...s introduced into a cell). For stable transfection, the DNA is introduced in such a way that it is maintained indefinitely. Today you will be transient
  10 KB (1,541 words) - 21:08, 28 July 2015
• 20.109(F11): Mod 1 Day 7 FACS analysis
  ... sample of this cell type from a complex mixture by using a FACS machine. In addition to purification, the FACS machine can count the number of cells th ...rged droplets, each containing a single cell. The stream of droplets pass in front of a laser beam, and the scattered light is analyzed by a series of f
  5 KB (840 words) - 21:08, 28 July 2015
• 20.109(F11): Mod 2 Day 2 Measuring system performance
  ...e output...then the cells might swim to the light rather than turn a color in response. ...nderstanding of the system we're working with is obviously needed. To look in the black box that senses light, we'll have to learn more about the two com
  21 KB (3,476 words) - 21:08, 28 July 2015
• 20.109(F11): Mod 2 Day 7 Readout of Protein
  ...ression of the light sensing protein, Cph8, and the mutant versions of it. In general, detection depends on which antibody you choose, and the quality of ...ize one epitope, while other antibodies, called polyclonal antibodies, are in fact antibody pools that recognize multiple epitopes.
  10 KB (1,644 words) - 21:08, 28 July 2015
• 20.109(F12): Mod 3 Day 1 Growth of phage materials
  ...rganisms themselves. Humans have a lot to learn from natures successes. In the upcoming weeks well use a virus that infects bacteria, namely the ba ...Esther Bullitt, Boston University School of Medicine), where the black bar in each image is 50 nm long. And what about the upper limit to the length of t
  18 KB (3,015 words) - 13:43, 29 July 2015
• 20.109(S13):Testing cell viability (Day3)
  ... to depletion by cells on the outer rim, even when at a high concentration in the bulk fluid. Finally, note that most cells require certain soluble and/o ...r in their cell permeance and nucleic acid affinity. Fluorescence emission in the green and red (left) and red alone (right) channels is shown for the sa
  15 KB (2,447 words) - 13:48, 29 July 2015
• 20.109(S13):Prepare expression system (Day4)
  ... your oh-so devoted teaching staff picked two colonies per mutant to grow in liquid culture. The XL1-Blue cell line, although it now carries the inverse ...galactoside). To reduce ‘leaky’ expression of the protein of interest (in our case, inverse pericam), the pLysS version of BL21(DE3) contains T7 lyso
  20 KB (3,344 words) - 13:48, 29 July 2015