Description
The CRISPRclean Repetitive Elements Depletion (Wheat) kit is designed to deplete wheat repetitive element sequences in just 4.5 hours with ~1 hours of hands-on time. This kit contains the reagents necessary to process the user’s adapter ligated DNA libraries through depletion. The protocol is suited for total DNA. The final product is a DNA library compatible with next-generation sequencing instruments.
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Description |
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Assay time |
~4.5 hours |
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Hands-on time |
~1 hour |
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Recommended library prep |
NEBNext Ultra II DNA Library Prep Kit |
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Ideal library fragment size |
750-900 bp |
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Input quantity for depletion |
~1 ng of adapter-ligated DNA library per sample |
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Depletion mechanism |
CRISPR Cas9 mediated |
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Multiplexing |
8 samples per depletion reaction |
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Species |
Wheat (tested on Chinese Spring and Jagger strains) |
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Designed to deplete |
Wheat repetitive elements |
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tested performance |
tested Chinese spring and jagger strains: • Chinese spring had a depletion rate of 69% of targeted region which translates to 54% removal of the wheat genome. • Jagger had a depletion rate of 54%, which translates to 42% removal of genome. |
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Compatibility |
Short read sequencing instruments such as Illumina® instruments |
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Samples per kit |
24 samples |
Kit contents, storage, and shelf life
The kit contains enough material to prepare 24 DNA libraries for Illumina® compatible sequencing. 8 wheat DNA libraries can be pooled in each depletion reaction. The shelf life of all reagents is 12 months from the date of manufacturing when stored properly.
The CRISPRclean Repetitive Element Depletion (Wheat) is made up of the following kit contents to be stored at the temperatures indicated in the table below:
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Depletion box Kit contents |
Storage temp |
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CRISPRclean Cas9 |
-20°C |
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CRISPRclean 10x Cas9 buffer |
-20°C |
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CRISPRclean Nuclease-free water |
-20°C |
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CRISPRclean RNase inhibitor |
-20°C |
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CRISPRclean 2X PCR mix |
-20°C |
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CRISPRclean P5 primer |
-20°C |
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CRISPRclean P7 primer |
-20°C |
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CRISPRclean Tris buffer |
-20°C |