Spider Cricket (B2)

Sample information

Picture
Photos by: Drew P
Location
Collection date 09/17/2023
Captive / Cultivated? Wild-caught
Group Edmund Burke School
Observations

The insect has spikes on its many legs. It didn’t have any fur or fuzziness on it and looks like it’s soft shelled. It has long antennas and has brown and golden colors.

Putative identification Arthropoda Hexapoda Insecta Orthoptera

Methods

Extraction kit DNeasy (Qiagen)
DNA extraction location Abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniOne
Buffer TBE
DNA stain GelGreen
Gel images
Protocol notes

DNA Extraction (10/04/23):

It was hard to crush up the body of the spider cricket. Even after crushing it up for awhile it wasn’t fully flat. The liquid was very chunky and almost stringy from the spider cricket.

I didn’t make any mistakes besides almost using the same pipette tip twice, but didn’t happen because my partner on the lab stopped me before I did it.

PCR (October 13, 2023):

No mistakes

Gel electrophoresis, Arthropod CO1 (October 18, 2023):

There was an outline of a band that appeared in this lane. It didn’t fully show up but you can defiantly still see it. There possibly might not have been enough DNA in the ladder. My DNA extraction possibly worked because it didn’t fully show up but something is still there.

Gel electrophoresis, Wolbachia, Arthropod PCR Day 2 (October 24, 2023):

Lane 1:  Ladder, Lane 2: Yellowjacket (B1), Lane 3: Spider Cricket (B2), Lane 4: Cicada (B3), Lane 5: Ladybug (B4), Lane 6: Positive Control, Lane 7: Negative Control, Lane 8: Water, Lane 9: Empty

  • No mistakes were made
  • All controls worked
  • Nothing is Wolbachia positive, besides the positive control.
  • Confidence level is high. I’m confident because the bands were not shown at all (besides the positive control) and it looked very clear meaning.

November 2023

*Arthropod CO1 was not sent to Penn State because the PCR Protocol had to be re-runned.

Results

Wolbachia presence No
Confidence level Medium
Explanation of confidence level

I’m confident that it’s not Wolbachia positive because no band showed up on the gel.

Wolbachia 16S sequence
Arthropod COI sequence
Summary The Orthoptera was found to be negative for Wolbachia.
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