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FluoroFun > 6. class single molecule fluor part 2 > Flashcards

6. class single molecule fluor part 2 Flashcards

1
Q

Single molecule enzymatics

A
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2
Q

Single molecule enzymatics dynamics

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3
Q

Time dependent kcat

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4
Q

The lipase B from the yeast candida antarctica

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5
Q

Non fluorescent substrate conversion

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6
Q

Long observation time possible

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7
Q

Several conformations are involved

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8
Q

How many conformations are involved?

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9
Q

The ‘lazy’ enzyme

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10
Q

Size of the fluorogenic substrate

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11
Q

Size of the substrate

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12
Q

Single molecule enzymatics

A
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13
Q

Single molecule enzymatics

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14
Q

Phospholipases

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15
Q

Phospholipase A1 studied

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16
Q

Single molecule enzymatics

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17
Q

Single molecule enzymatics

first step

A
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18
Q

Single molecule enzymatics?

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19
Q

Water soluble perylenediimide dye (PDI)

A
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20
Q

Dual color excitation

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21
Q

Active PLA1 – High concentration

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22
Q

Active PLA1 – Low concentration

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23
Q

Active PLA1 vs inactive

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24
Q

Single enzyme trajectories

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25
Q

Image analysis

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26
Q

hetero vs Homogeneous track

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27
Q

Cummulative distribution functions

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28
Q

Enzyme mobility

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29
Q

Model of mode of operation

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30
Q

Getting closer to cell membranes….

31
Q

Getting closer to nature….

32
Q

Hydrolysis of giant unilamellar vesicles (GUV’s)

33
Q

cytoplasma & er

34
Q

YFP-PLCβ1 – Ca2+ response

35
Q

eGFP-PLCβ1 – Ca2+ response + SPT

36
Q

DNA sequencing & the human genome project

37
Q

DNA sequencing

38
Q

DNA-sequencing history

39
Q

1 st gen: Sanger

40
Q

1 st gen: Sanger method

41
Q

1 st gen: Automated Sanger Method

42
Q

1 st gen: Automated Sanger Method

43
Q

1 st gen: Automated Sanger Method

44
Q

2 nd gen: 454 sequencing

45
Q

2 nd gen: 454 sequencing (Roche®)

46
Q

2 nd gen: 454 sequencing (Roche®)

47
Q

2 nd gen: 454 sequencing (Roche®)

48
Q

2 nd gen: Illumina® (Solexa) sequencing

49
Q

2 nd gen: Illumina® (Solexa) sequencing

50
Q

reversible terminators

51
Q

comparison of next generation sequencing platforms

52
Q

Single molecule DNA Sequencing

53
Q

Microcapillary-based SM DNA sequencing (M.Sauer et al. Heidelberg)

54
Q

Microcapillary-based SM DNA sequencing

55
Q

Microcapillary-based SM DNA sequencing

56
Q

Flow-cytometry based DNA sequencing

57
Q

cleavage vs control

58
Q

DNA-sequencing: the Helicos-approach

step 1

59
Q

DNA-sequencing: the Helicos-approach

step 2

60
Q

DNA-sequencing: the Helicos-approach

step 3

61
Q

DNA-sequencing: the Helicos-approach

step 4

62
Q

DNA-sequencing: the Helicos-approach

step 5

63
Q

DNA-sequencing: the Helicos-approach

step 6

64
Q

DNA-sequencing: the Helicos-approach

step 7

65
Q

DNA-sequencing: the Helicos-approach

step 8

66
Q

DNA-sequencing: the Helicos-approach

step 9

67
Q

helicos tool

68
Q

correlogram

69
Q

DNA target template

70
Q

HELICOS & FRET

71
Q

Pacific biosciences approach:

72
Q

metal film

73
Q

Pore-based sequencing

FluoroFun (9 decks)

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