Protocols and Recipes
Below are protocols and recipes used in Blattner lab.
Microarray
E. coli
Total RNA Labeling for High Density Oligonucleotide Array [PDF]
Labeling E. coli total RNA for high density oligonuceotide array
like Affymetrix E. coli antisense genome array. [updated/corrected
05/30/2007]
E. coli
Total RNA Isolation [PDF]
Stablize E. coli RNA with Qiagen Bacteria RNA Protect Reagent and
isolate total RNA with Epicentre MasterPure RNA Purification Kit. [updated
07/31/2003]
E. coli
Total RNA Labeling for Spotted Microarray [PDF]
Label E. coli Total RNA with CyDye-dUTP directly using Invitrogen
Superscript II reverse transcriptase. [updated/corrected
05/30/2007]
E. coli Genomic
DNA Labeling [PDF]
Label fragmented E. coli genomic DNA with CyDye-dUTP directly using
Klenow Fragment. [updated 02/10/2002]
Spotted
array Hybridization [PDF]
Hybridize CyDye-dUTP labeled probes to E. coli microarray printed
on poly-lysine coated slide (provided by GEC) using Sigma ArrayHyb Buffer.
[updated 02/10/2002]
Mutagenesis
Tn5 Mutagenesis
[PDF]
Mutate E. coli K-12 genes with Tn5 Transposon. [updated 05/11/2002]
Media
These are our protocols for preparing Neidhardt's MOPS-based defined
media. These media are used for the global transcription analyses, and
are now available commercially
from TekNova Inc.
EZ Rich Defined Medium
[PDF]
MOPS Minimal Medium
[PDF]
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