Tumor vs normal on Single Clones 10-11-12

2015-01-13

azim58 - Tumor vs normal on Single Clones 10-11-12

10-11-12

For this experiment I plan to print protein lysate from single colonies
(see 8-26-12 Plasmids for verification) onto nitrocellulose arrays (see
Tumor Lysate Nitrocellulose Print 10-12-12). These arrays will also have
plain e coli lysate printed onto the array as a negative control. The
arrays will be probed with naive sera and tumor sera. I will apply 3
dilutions of sera onto these slides: 1:2000, 1:1000, and 1:500. With this
experiment I would like to see if the results are repeateable from the
last (Tumor vs normal on Single Clones 9-21-12), and I would also like to
see if an even more dilute aliquot of sera brings out the intensity of
tumor against naive even more. I will use 3 slides for each dilution.

Print will take place on 10-12-12
experiment performed 10-13-12
Slides were scanned 10-14-12
raw data here
"S:\Research\Cancer_Eradication\Users\kwhittem\kwhittem\Raw Data Often
Originally on Research Drive\2012\10-14-12 nv-tmr-lst\nv-tmr-lst 10-14-12"

analysis here
"C:\kurt\storage\CIM Research Folder\DR\2012\10-21-12\lysate array\naive
tumor lysate 10-21-12.xlsx"
volcano plots here
"C:\kurt\storage\CIM Research Folder\DR\2012\10-21-12\lysate
array\volcano plots 10-21-12.svg"

This experiment did show the general trend I've been seeing in other
experiments. Generally, the more the dilute the sample (up to a point)
the more intensity the tumor has than the naive. In this experiment, I
think I scanned with too high of a gain though. I scanned at a higher
gain than usual, which I think resulted in more noise. There was less of
a difference between the spots in this experiment than in some other
experiments. I am considering rescanning the slides with a high laser
power but low gain, but the slides may be too old now to still obtain
good results.

===========================================================================
I rescanned the slides at 100l10g since I thought the gain was too high
on the last scan. Note that the slides may have been sitting too long
before I did this rescan.

raw data here
"S:\Research\Cancer_Eradication\Users\kwhittem\kwhittem\Raw Data Often
Originally on Research Drive\2012\10-22-12\nv-tmr-lst 10-22-12"

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