ProtocolsLeafhoppers < Lab

---+ 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_ %BR%
_Curtara insularis_ %BR%
_Graminella nigrinota_ %BR%

These all happily reproduce on barley, which is easy to grow and maintain in lab environments. 

---+++ Protocols and Primers

Leafhopper COI Lep Primers:%BR%
LepF: 5'-ATTCAACCAATCATAAAGATATTGG-3' %BR%
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.

Barrick Lab > ProtocolList > ProtocolsLeafhoppers

Topic revision: r3 - 2021-11-03 - 20:09:36 - Main.SarahBialik