Biomod/2014/HKBUteam/experimentalresults: Difference between revisions
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<b>Mass-spectrometry</b> | <b>Mass-spectrometry</b> | ||
1. Control group | |||
1. Control group one: MS-Single stranded DNA A2<br> | |||
[[Image:Mass_Spectrometry_Control_Single-stranded_DNA_A2.png|500px|For MS-single stranded DNA A2 control ]] | [[Image:Mass_Spectrometry_Control_Single-stranded_DNA_A2.png|500px|For MS-single stranded DNA A2 control ]] | ||
2. Control group | 2. Control group two: MS-Single stranded DNA A3<br> | ||
[[Image:Mass_spectrometry_Single-stranded_DNA_A3_.png|500px|For MS-single stranded DNA A3 control ]] | [[Image:Mass_spectrometry_Single-stranded_DNA_A3_.png|500px|For MS-single stranded DNA A3 control ]] | ||
3. conjugated group<br> | |||
3. | |||
[[Image:Exp1.2.jpg|720px|thumb|alt=Example alt text|For MS-single stranded DNA A2 ,a minor peak at 10120 shows the conjugation of A2 and chlorambrucil.<br>For MS-single stranded DNA A3 , a minor peak at 7043 shows the conjugation of A3 and folic acid.<br>For MS-single stranded DNA A6 , a peak at 11171 shows the conjugation of A6 and carbon nanodots<br>]] | [[Image:Exp1.2.jpg|720px|thumb|alt=Example alt text|For MS-single stranded DNA A2 ,a minor peak at 10120 shows the conjugation of A2 and chlorambrucil.<br>For MS-single stranded DNA A3 , a minor peak at 7043 shows the conjugation of A3 and folic acid.<br>For MS-single stranded DNA A6 , a peak at 11171 shows the conjugation of A6 and carbon nanodots<br>]] | ||
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Gel photo : | Gel photo : | ||
[[Image:expresultSection1_4.jpg|400px| | [[Image:expresultSection1_4.jpg|400px|]] <br> | ||
GeneRuler DNA Ladders, Samples, Modified Primers A2, A3 <br> | |||
<h4>Nanodrop Reading</h4> | <h4>Nanodrop Reading</h4> |
Revision as of 10:46, 25 October 2014
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1st Trial
Conjugation condition
Single strand DNA, A2 and A3 conjugated with chlorambrucil and folic acid respectively in ratio 1:1. Mixtures were vortexed at room temperature for 4 hrs.
In-Complete route
Folic acid and chlorambrucil were first conjugated and folded, carbon nanodots were added afterwards at ration 1:1.
Mass-spectrometry
1. Control group one: MS-Single stranded DNA A2
2. Control group two: MS-Single stranded DNA A3
3. conjugated group
DNA extraction after folding of DNA origami
Gel photo :
GeneRuler DNA Ladders, Samples, Modified Primers A2, A3
Nanodrop Reading
DNA concentration : 0.8 ng/ul
260/280 : 1.34
260/230 : 0.86
Confocal Microscopy
Hep G2 Treatment
Hep G2 Control
Excitation wave lengths: 405nm
Emission wave lengths: 460 – 560nm
2nd Trial
Conjugation condition
Single strand DNA, A2 and A3 conjugated with chlorambrucil and folic acid respectively in ratio 1:1. Mixtures were vortexed at room temperature for 4 hrs.
In-Complete route
Folic acid and chlorambrucil were first conjugated and folded, carbon nanodots were added afterwards at ration 1:1.
Mass-spectrometry
1. Control group 1 & 2 refer to Trial 1
2. Conjugated group
For MS-single stranded DNA A2 , a minor peak at 10120 shows the conjugation of A2 and chlorambrucil.
For MS-single stranded DNA A3 , a minor peak at 7043 shows the conjugation of A3 and folic acid.
DNA extraction after folding of DNA origami
Nanodrop Reading
DNA concentration : 2.3 ng/ul
260/280 : 1.28
260/230 : 0.52
Confocal Microscopy
Hep G2 Treatment
Hep G2 Control
Excitation wave lengths: 405nm
Emission wave lengths: 460 – 560nm
3rd Trial
Conjugation condition
Single strand DNA, A2, A3 and A6 conjugated with chlorambrucil, folic acid and carbon nanodots respectively in ratio 1:10.
Mixtures were vortexed at 37 for 4 hrs.
Complete route
All substituent was conjugated and folded simultaneously
Mass-spectrometry
1. Control group 1 & 2 refer to Trial 1
2. Conjugated group
DNA extraction after folding of DNA origami
Gel photo :
Nanodrop Reading
DNA concentration : 5.5 ng/ul
260/280 : 2.65
260/230 : 1.12
Confocal Microscopy
Hep G2 Treatment
Hep G2 Control
HK-1 Treatment
HK-1 Control
Excitation : 405nm
Emission : 460 – 560nm
Emission Spectrum
Hep G2 HK-1
AFM photo
Discussion:
The Atomic force microscopy image has a low resolution and the structure of the functionalized DNA origami
can hardly be identified which may due to the following reasons:
- The DNA origami samples is not dried up completely which increase the adhesive force of the AFM instrument
- The DNA origami samples was clustered on the mica platform
- The scan size of AFM is restricted from 1 to 5 nm so many area attached with the DNA was not examined properly