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Protocols and Recipes
Below are protocols and recipes currently 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 05/30/2007. [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 K12 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