Difference: ProtocolsLeafhoppers (1 vs. 3)
Revision 32021-11-03 - SarahBialik
Leafhopper Care and ProtocolsRearing and Caring for Leafhopper Species | ||||||||
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| < < | Various leafhoppers species are kept in BugDorms and maintained on various plants (barley, Virginia creeper). | |||||||
| > > | Leafhopper species are kept in BugDorms (BugDorm-4F3030 and 4F2222). We have the following colonies reliably growing in lab: | |||||||
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| > > | Macrosteles quadrilineatus Curtara insularis Graminella nigrinota These all happily reproduce on barley, which is easy to grow and maintain in lab environments. | |||||||
Protocols and Primers | ||||||||
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| < < | Leafhopper Primers: | |||||||
| > > | Leafhopper COI Lep Primers: | |||||||
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| > > | LepF: 5'-ATTCAACCAATCATAAAGATATTGG-3' LepR: 5'-TAAACTTCTGGATGTCCAAAAAATCA-3' | |||||||
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| < < | Barcoding uses a gDNA extraction kit and PCR using above primers. | |||||||
| > > | We use the Qiagen DNeasy Blood and Tissue Kit to extract gDNA from each individual leafhopper. We then perform PCR using the OneTaq polymerase and follow the standard protocol provided by NEB. We use Lep primers (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1327734/) which we also use for IDing aphids. We run a gel to confirm a band around 700bp and then purify the PCR product with the Zymo Clean and Concentrate kit. It is submitted for Sanger Sequencing using both the forward and reverse primers and aligned in Benchling. When the full COI sequence is obtained, we then use the BOLD database to search for the leafhoppers identity. | |||||||
Revision 22020-12-08 - SarahBialik
Revision 12020-12-04 - SarahBialik
Leafhopper Care and ProtocolsRearing and Caring for Leafhopper SpeciesBarley is the best.Protocols and PrimersI love leafhoppers. |
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