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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

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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

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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

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18
Q

Single molecule enzymatics?

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19
Q

Water soluble perylenediimide dye (PDI)

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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….

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31
Q

Getting closer to nature….

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32
Q

Hydrolysis of giant unilamellar vesicles (GUV’s)

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33
Q

cytoplasma & er

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34
Q

YFP-PLCβ1 – Ca2+ response

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35
Q

eGFP-PLCβ1 – Ca2+ response + SPT

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36
Q

DNA sequencing & the human genome project

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37
Q

DNA sequencing

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38
Q

DNA-sequencing history

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39
Q

1 st gen: Sanger

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40
Q

1 st gen: Sanger method

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41
Q

1 st gen: Automated Sanger Method

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42
Q

1 st gen: Automated Sanger Method

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43
Q

1 st gen: Automated Sanger Method

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44
Q

2 nd gen: 454 sequencing

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45
Q

2 nd gen: 454 sequencing (Roche®)

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46
Q

2 nd gen: 454 sequencing (Roche®)

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47
Q

2 nd gen: 454 sequencing (Roche®)

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48
Q

2 nd gen: Illumina® (Solexa) sequencing

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49
Q

2 nd gen: Illumina® (Solexa) sequencing

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50
Q

reversible terminators

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51
Q

comparison of next generation sequencing platforms

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52
Q

Single molecule DNA Sequencing

A
53
Q

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

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54
Q

Microcapillary-based SM DNA sequencing

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55
Q

Microcapillary-based SM DNA sequencing

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56
Q

Flow-cytometry based DNA sequencing

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57
Q

cleavage vs control

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58
Q

DNA-sequencing: the Helicos-approach

step 1

A
59
Q

DNA-sequencing: the Helicos-approach

step 2

A
60
Q

DNA-sequencing: the Helicos-approach

step 3

A
61
Q

DNA-sequencing: the Helicos-approach

step 4

A
62
Q

DNA-sequencing: the Helicos-approach

step 5

A
63
Q

DNA-sequencing: the Helicos-approach

step 6

A
64
Q

DNA-sequencing: the Helicos-approach

step 7

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65
Q

DNA-sequencing: the Helicos-approach

step 8

A
66
Q

DNA-sequencing: the Helicos-approach

step 9

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67
Q

helicos tool

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68
Q

correlogram

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69
Q

DNA target template

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70
Q

HELICOS & FRET

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71
Q

Pacific biosciences approach:

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72
Q

metal film

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73
Q

Pore-based sequencing

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FluoroFun (9 decks)

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