Difference: ProtocolsLeafhoppers (1 vs. 3)

Revision 32021-11-03 - SarahBialik

 
META TOPICPARENT name="ProtocolList"

Leafhopper Care and Protocols

Rearing and Caring for Leafhopper Species

Changed:
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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:
 
Added:
>
>		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

Changed:
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<		Leafhopper Primers:
>
>		Leafhopper COI Lep Primers:
Added:
>
>		LepF: 5'-ATTCAACCAATCATAAAGATATTGG-3'
LepR: 5'-TAAACTTCTGGATGTCCAAAAAATCA-3'
 
Changed:
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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

 
META TOPICPARENT name="ProtocolList"

Leafhopper Care and Protocols

Rearing and Caring for Leafhopper Species

Changed:
<
<		Barley is the best.
>
>		Various leafhoppers species are kept in BugDorms and maintained on various plants (barley, Virginia creeper).
 

Protocols and Primers

Changed:
<
<		I love leafhoppers.
>
>		Leafhopper Primers:
Added:
>
>		 Barcoding uses a gDNA extraction kit and PCR using above primers.
 

Revision 12020-12-04 - SarahBialik

 
META TOPICPARENT name="ProtocolList"

Leafhopper Care and Protocols

Rearing and Caring for Leafhopper Species

Barley is the best.

Protocols and Primers

I love leafhoppers.

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