TILLING (Targetting Induced Local Lesions In Genomes) Technology for
Plant Functional Genomics
R. Soner SİLME, M. İlhan ÇAĞIRGAN http://www.nobelonline.net/UserFiles/File/PDF.pdf

Journal of Applied Biological Sciences 1 (1): 77-80, 2007

TILLING used to detect gene lesions obtained through chemical mutagenesis

Isolate DNA, PCR amplify using wild type primers.

Mutant and WT DNA will hybridise, but areas of difference will cause the DNA to loop out.

Add S1 nuclease, which will cleave mismatched strands.

Fluorescent end labelling will make it easy to identify mutant strands when run on a gel (look for RFLPs)




H2 directed evolution experiment:
Mutate cells
Transfer to microplate with cell lysis chemicals, universal pH indicator and an acid
H2ase activity will use up protons and result in colour change?

Another idea:
Transform E coli with combinatorially mutagenised H2ase. Grow in LB and then plate out on acid agar (pH 4.3). H2ase will convert protons to H2 and have protective effect?