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"