Editing
Comparison of Staining Dyes
(section)
Jump to navigation
Jump to search
Warning:
You are not logged in. Your IP address will be publicly visible if you make any edits. If you
log in
or
create an account
, your edits will be attributed to your username, along with other benefits.
Anti-spam check. Do
not
fill this in!
=4/16/10= ==Purpose== To establish the best dye to use for any subsequent experiments in order to maintain consistency in results. ==Methods== 3 dyes were tested: Dapi, SYBR Green, and YOYO. Each dye was diluted to 2X concentration. Sample tubes 1 and 2 were combined, filtered through a 5um filter, and used as a standardized sample. For each dye: - Use a clean 0.2mL tube - Add 7uL sample + 3uL 2X dye - Incubate for 10 minutes in the dark - Add 10uL water - Vortex and spin down Each sample was plated and imaged under a fluorescence microscope using 10X magnification. ==Results== [[image: Dapi_10X_Comparison.jpg]] ''Image 1: 2X Dapi'' [[image: SYBR Green_10X_Comparison.jpg]] ''Image 2: 2X SYBR Green'' [[image: YOYO_10X_Comparison.jpg]] ''Image 3: 2X YOYO'' ==Discussion== (4/16/10) Even though the images appeared to be dirty, SYBR Green showed the clearest image, so we decided to stick with SYBR Green for the time being. However, we were unsure whether or not glycerol in the sample affected the image quality. To help reassess the sample, we performed a follow-up experiment. (4/22/10) Because of SYBR Green's rather fast photo-degradation, we have decided to try and standardize our samples with 2X Dapi staining. The issue with Dapi is that it can produce saturated images, so the concentration must be carefully considered. ==Follow-up== ===Methods=== Two slides were extensively cleaned: washed down with isopropanol, wiped with a Kimwipe, washed again with isopropanol, and air-dried in the fume hood. To one slide a 20% glycerol mixture was added, with 2X SYBR Green as the dye. To the other slide, 7uL sample + 3uL 2X SYBR Green was added. ===Results=== [[image: 20%Glycerol_10X.jpg]] ''Image 1: 20% Glycerol strained with 2X SYBR Green'' [[image: Sample_cleaned_10X.jpg]] ''Image 2: Sample stained with SYBR Green'' ===Discussion=== Unexpectedly, both the glycerol control and the samples had masses show up under the microscope. This is odd because the slides were thoroughly cleaned and the glycerol did not have chromosomes added to it. This suggests that glycerol does in fact bind to the SYBR Green dye.
Summary:
Please note that all contributions to ZhangLabWiki may be edited, altered, or removed by other contributors. If you do not want your writing to be edited mercilessly, then do not submit it here.
You are also promising us that you wrote this yourself, or copied it from a public domain or similar free resource (see
ZhangLabWiki:Copyrights
for details).
Do not submit copyrighted work without permission!
Cancel
Editing help
(opens in new window)
Navigation menu
Personal tools
Not logged in
Talk
Contributions
Create account
Log in
Namespaces
Page
Discussion
English
Views
Read
Edit
View history
More
Search
Navigation
Main Page
Current events
Recent changes
Random page
Investigators
Matt Cai
Song Chen
Eric Chu
Dinh Diep
Elizabeth Duong
Shicheng Guo
Alan Fung
Daniel Jacobsen
Blue Lake
Huy Lam
Alice Li
Andrew Richards
Brandon Sos
Chris Wei
Yan Wu
Kun Zhang
Tools
What links here
Related changes
Special pages
Page information