Printing Tumor cDNA Library

2015-01-13

azim58 - Printing Tumor cDNA Library

042811 Printing cDNA Library onto Array

Cell Lysate Test Prints

Initial Notes (wuala link)
Initial Notes 042811
Printing Actual Library 010912
Printing 2-17-12_16E3 Spot

Planning -> 042811 -> 050211 -> (Results) 5-16-11

Planning ->062411 (2nd Library on Array Experiment)
Protocols
Standard Bacteria Lysis Protocol of Lab (from Alan):
http://www.wuala.com/azim58/wuala/CIM%20Research%20Folder/kwhittem/Records%
20in%20CIM%20Folder/Dated%20Records/2011/4-28-11/Bacteria%20Lysis%20Protoco
l%20from%20Alan.pdf/

Diagram of Protocol for Plate Format
Version 1: 4-28-11
Version 2: 5-24-11
Version 3: 6-20-11 (this version uses the correct amount of lysozyme)
Slides
Slides Types

Map of Experiments ->
5-3-11 Transform Library and aliquot into plate for overnight culture ->
Perform colony PCR to see if diversity is maintained
5-4-11 Transformation efficiencies
5-5-11 Plating of overnight plate cultures
5-6-11 Colony PCR of Tumor Library Culture
5-9-11 Gel of colony PCR
5-10-11 Extract Tumor Library Proteins
5-13-11 Initial Tumor Library Array Experiment
5-16-11 Results of Initial Tumor Library Array Experiment

5-17-11 Western Blot to Check for SMC1fs Protein
5-18-11 2nd Half of Western Blot
5-19-11 Western Blot Results (SMC1fs Protein Membrane) (with DH10B cells)

C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\5-19-11

5-24-11 Extract proteins from BL21 containing pET32b and SMC1fs (see
notebook)
5-31-11 Western Blot Results (SMC1fs Protein Membrane) (with BL21 cells)

C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\5-31-11\SMC1fs Western Blot

6-2-11 Protein Extraction from BL21 cells (no plasmid, pET32b, and
pGEX-SMC1fs-27mer)
6-3-11 Western Blot to Detect Extracted Protein
6-6-11 Western Blot Results

C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\6-6-11\Western Blot and Coomassie Results

6-(8-9)-11 Extraction of Protein with lysozyme and sonication

6-13-11 Coomassie to Identify 14 kD band

"C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\6-13-11\Coomassie Annotated 6-13-11.svg"

6-(14-16)-11 Test of anti-rabbit for 14 kD Band
Autofluorescence measured here

"C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\6-14-11\Scan for 14 kd autofluorescence\Scan for 14 kD
autofluorescence annotated.svg"

test for secondary binding here
C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\6-17-11\Test of Secondary for 14 kD Band

6-20-11 Protein Expression w/less lysozyme
6-(21-22)-11 Western Blot to Check for 14 kD Band

"C:\kurt\storage\CIM Research Folder\kwhittem\Records in CIM Folder\Dated
Records\2011\6-23-11\Western Blot for 14 kD Band 6-23-11\Annotated
Files\Western Blot for 14 kD Band 6-23-11.svg"

6-(24-(7-5))-11 Printing cDNA Library 2nd Attempt

7-8-11 Printing cDNA Library 3rd Attempt
7-14-11 Cell Lysate Plate Preparation
Made denatured plate too
7/12/11 Cell Lysate Slide Results 3 (CodeLink and Aminosilane)
7/18/11 Cell Lysate Printing 4 (Non-denatured and denatured)
8/4/11 Cell Lysate Printing 5 (hepes and glycerol test)
11/23/11 Cell lysate printing onto ArrayIt Slides 11-23-11

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