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Figure 5 | Plant Methods

Figure 5

From: NEATTILL: A simplified procedure for nucleic acid extraction from arrayed tissue for TILLING and other high-throughput reverse genetic applications

Figure 5

Identification of cleaved fragments of hp1 mutant after CEL I digestion in Licor 4300 DNA Analyzer. Positive control DNA bearing hp1 mutation was laced with eight-fold pooled DNA which was isolated by NEATTILL procedure as described in the text. The expected position of the CEL I cleaved fragments are 718 bp (bottom left arrow) in 700 channel and 183 bp (right arrow) in 800 channel which were detected in all replicates. These cut fragments add up to 901 bp (top left arrow) (total amplicon length of hp1 gene). M on right hand side of the two images shows 50-700 bp size standards. The fragments detected specially in the lower portion of either channel images represent the background fragments usually observed in TILLING runs. However these can be distinguished from the CEL I cleaved fragments as these fragments do not show the complementary fragment in the either channel, a characteristic of CEL I cleaved fragments.

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Plant Methods

ISSN: 1746-4811