Leafhopper Care and Protocols

Rearing and Caring for Leafhopper Species

Leafhopper species are kept in BugDorms (BugDorm-4F3030 and 4F2222). We have the following colonies reliably growing in lab:

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

Leafhopper COI Lep Primers:
LepF: 5'-ATTCAACCAATCATAAAGATATTGG-3'
LepR: 5'-TAAACTTCTGGATGTCCAAAAAATCA-3'

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.

Edit | Attach | Watch | Print version | History: r3 < r2 < r1 | Backlinks | Raw View | More topic actions

 Barrick Lab  >  ProtocolList  >  ProtocolsLeafhoppers

Contributors to this topic Edit topic SarahBialik
Topic revision: r3 - 2021-11-03 - 20:09:36 - Main.SarahBialik
 
This site is powered by the TWiki collaboration platform Powered by Perl This site is powered by the TWiki collaboration platformCopyright ©2022 Barrick Lab contributing authors. Ideas, requests, problems? Send feedback