Sample information

-Black beetle with a horizontal brown strip across the back where the Neck attaches to the Body.

Methods

1.) I got my DNA through the protocol found in lab 2: DNA extraction. However, when I did my cell Lysis step, I left the test tube in the heat for 30 minutes, vortexing every 10 minutes.
2.) Next, I followed the protocol found in lab 3: PCR with Athropod primers. 3/4/25
3.) followed Lab:4 pt 1 Gel electrophoresis w/ arthropod primers See gel image above.
4.) Followed lab 3 part 2: PCR round 2
For this step, I used the NEB OneTaq hot start as my Taq polymerase. I also ran my Annealing temp at 55c on 3/31/25.
5.) On 4/1/25 I ran a single PCR reaction on my DNA. I used Lab 4 part 2 from the Wolbachia project. I used a gel electrophoresis system to move my DNA and check for Wolbachia DNA. I used a TAE buffer and a SYBR safe DNA stain. The polymerase used was: New England Biolabs OneTaq Hot start quick-load 2x MM w/ Standard buffer (product # M0488s). The DNA ladder I used was: New England Biolabs 1 kb plus DNA ladder for safe stains (product #N0559s). I only did one PCR on 3/31/25 using the NEB OneTaq hot start as my Taq polymerase. I also ran my Annealing temp at 55c.

Final:
The leader Beetle was Wolbachia +
With a high confidence level
I am highly confident because there were no smears throughout my entire process. My controls also had or didn’t have lines when necessary. Such as when looking for DNA in water none was present and while testing my positive arthropod control, arthropod was present.

I am highly confident because there were no smears throughout my entire process. My controls also had or didn’t have lines when necessary. Such as when looking for DNA in water none was present and while testing my positive arthropod control, arthropod was present.