Biomod/2011/TUM/TNT/LabbookA/Preparing samples for transmission electron microscope: Difference between revisions

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D2: BM2 with 0.74 µM ethidium bromide (corresponding to one molecule every 21bp) <br>
D2: BM2 with 0.74 µM ethidium bromide (corresponding to one molecule every 21bp) <br>
D3: BM2 without any DNA binders <br>
D3: BM2 without any DNA binders <br>
<br>
[http://openwetware.org/wiki/Biomod/2011/TUM/TNT/LabbookA/2011/10/14 14th October] <br>
BM2 dilution: 1:25 <br>
D4: BM2 with 432 nM DAPI (corresponding to one molecule every 7bp) <br>
D5: BM2 with 144 nM DAPI (corresponding to one molecule every 21bp) <br>
<br>
<br>

Revision as of 04:12, 14 October 2011

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       TUM NanU - Labbook

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preparing samples for transmission electron microscope

glow-discharge grids at 30mA for 45s
add 1µl 5M NaOH to 200µl 2% uranyl acetate staining solution
hide solution from light
vortex 1minute
centrifuge for 2.5 minutes at maximum speed
apply 5µl sample on grid
incubate for 3 minutes
remove liquid from grid without touching its sourface
lay grid on 20µl staining solution, then remove stain immediatly from grid
lay grid on another drop of 20µl staining solution
incubate for 40s seconds
remove liquid from grid
let grid dry for at least 15 minutes

grid positions


BIOMOD grid box

30th August
A2: 2DH3ML_BM1
A3: 2DH3ML_BM2
A4: 5DH3ML_BM1
A5: 5DH3ML_BM2

16th September
B2: BM2 control without DNA-binders
B3: BM2 with 10µM spermine
B4: BM2 with 30µM spermine
B5: BM2 with 100µM spermine
C2: BM2 with 25µM ethidium bromide
C3: BM2 with 75µM ethidium bromide
C4: BM2 with 250µM ethidium bromide

28th September
C5: BM2 control without DNA-binders
D1: BM2 with 6µM EtBr
D2: BM2 with 12µM EtBr
D3: BM2 with 2.4µM spermine
D4: BM2 with 4.8µM spermine
E1: BM2 with 0.9nM DAPI
E2: BM2 with 1.8nM DAPI

BIOMOD 2 grid box

5th October
A1: BM2 with 1.34 µM spermine
A2: BM2 with 1.34 µM spermine
A3: BM2 with 1.34 µM spermine
A4: BM2 with 0.42 µM spermine
A5: BM2 with 0.42 µM spermine
B1: BM2 with 0.42 µM spermine
samples A1, A2 and A3 correspond to one spermine molecule in average every 7bp, samples A4, A5 and B1 correspond to one molecule spermine in average every 21bp

10th October
BM2 dilution: 1:40
B2: BM2 with 2.27 µM ethidium bromide
B3: BM2 with 2.27 µM ethidium bromide
B4: BM2 with 2.27 µM ethidium bromide
B5: BM2 with 0.69 µM ethidium bromide
C1: BM2 with 0.69 µM ethidium bromide
C2: BM2 with 0.69 µM ethidium bromide
C3: BM2 without DNA binders
C4: BM2 without DNA binders
C5: BM2 without DNA binders
samples B2, B3 and B4 correspond to one ethidium bromide molecule in average every 7bp, samples B5, C1 and C2 correspond to one molecule ethidium bromide in average every 21bp

11th October
BM2 dilution: 1:25
D1: BM2 with 2.4 µM ethidium bromide (corresponding to one molecule every 7bp)
D2: BM2 with 0.74 µM ethidium bromide (corresponding to one molecule every 21bp)
D3: BM2 without any DNA binders

14th October
BM2 dilution: 1:25
D4: BM2 with 432 nM DAPI (corresponding to one molecule every 7bp)
D5: BM2 with 144 nM DAPI (corresponding to one molecule every 21bp)

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