Sam:LabNotes/Microbiome-new/2010-6-9 exp1: Difference between revisions
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>Sam Chiang (Created page with '='''VA-hospital sorting on Alex-FISH E.coli (EtOH-fixed) - with 1000cell pre-run'''= ==Background== *The new FISH-labeled E.coli was performed with only 30min hybridization (Mad…') |
>Sam Chiang |
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='''VA-hospital sorting on Alex-FISH E.coli (EtOH-fixed) - | ='''VA-hospital sorting on Alex-FISH E.coli (EtOH-fixed) - 30min-hybridization samples'''= | ||
==Background== | ==Background== | ||
*The new FISH-labeled E.coli was performed with only 30min hybridization (Made on 06-07-'10) | *The new FISH-labeled E.coli was performed with only '''30min hybridization''' (Made on 06-07-'10) | ||
*For this sorting, I pre-run a 1000cell sorting to cover full plate before the real sample sorting | *For this sorting, I pre-run a 1000cell sorting to cover full plate before the real sample sorting | ||
Line 24: | Line 24: | ||
A 100 100 100 100 100 100 100 100 100 100 100 100 | A 100 100 100 100 100 100 100 100 100 100 100 100 | ||
-------------------------------------------------- | -------------------------------------------------- | ||
==Results== | ==Results== | ||
[[File:sam060910-Ecoli sorting-new.jpg|800px]] | |||
==Discussion== | ==Discussion== | ||
*The 30min hybridization showed similar results as 90min hybridization protocol. | |||
*'''There still have two peaks showing when the sorting number is increased'''. | |||
*The background is very low in this result compared to the previous results (03-30-'10). '''I am not sure if the short-hybridization helps to reduce the negative-labeling'''. | |||
**http://genome-tech.ucsd.edu/LabNotes/index.php/Sam:LabNotes/Microbiome-new/2010-3-30 |
Latest revision as of 21:45, 18 June 2010
VA-hospital sorting on Alex-FISH E.coli (EtOH-fixed) - 30min-hybridization samples[edit]
Background[edit]
- The new FISH-labeled E.coli was performed with only 30min hybridization (Made on 06-07-'10)
- For this sorting, I pre-run a 1000cell sorting to cover full plate before the real sample sorting
Procedure[edit]
- Add 900uL 1XPBS into the sample (100uL) in the amber tube.
- Material need for sorting:
- 8-well PCR tube stirps (on 96-well rack). Filled up 3uL 1X PBS in PCR tubes. UV-treatment for 15min.
- UV clean sealing film (Bio-rad)
- UV clean zip bag for transporting sample + rack
Real sorting arrangement 1 2 3 4 5 6 7 8 9 10 11 12 - Strip# (Strips are in vertical arrangement) ------------------------------------------------- H 0 0 0 0 0 0 0 0 0 0 0 0 G 1 1 1 1 1 1 1 1 1 1 1 1 F 1 1 1 1 1 1 1 1 1 1 1 1 E 1 1 1 1 1 1 1 1 1 1 1 1 D 1 1 1 1 1 1 1 1 1 1 1 1 C 1 1 1 1 1 1 1 1 1 1 1 1 B 10 10 10 10 10 10 10 10 10 10 10 10 A 100 100 100 100 100 100 100 100 100 100 100 100 --------------------------------------------------
Results[edit]
File:Sam060910-Ecoli sorting-new.jpg
Discussion[edit]
- The 30min hybridization showed similar results as 90min hybridization protocol.
- There still have two peaks showing when the sorting number is increased.
- The background is very low in this result compared to the previous results (03-30-'10). I am not sure if the short-hybridization helps to reduce the negative-labeling.