Sample information

Tan colored insect that is approximately 1 inch in length.  It has antenna, legs and they hold their wings flat over their backs since they no longer fly.  These are typically sold in local pet stores to feed pets like lizards.  I purchased this cricket at PetCo in Pittsfield, MA.

Methods

Taq Polymerase (New England Biolabs One Taq Hot Start) used, Start Quick-Load 2x MM w/ Standard Buffer (product #M0488S.

DNA Ladder Used: New England Biolabs 1kb Plus DNA Ladder for Safe Stains (product #N0559S)

Annealing temperatures 1 cycle 30 sec @ 94 C, 30 cycles 30 sec @ 94 C and 45 sec @ 49 C and 60 Sec @ 68 C, 1 cycle 5 min @ 68 C.  Arthropod PCR reaction completed on 3/13/25, annealing temperature 49C.  Wolbachia (single) PCR reaction completed on 4/3/25, annealing temperature 55C.

The arthropod sample had a lot of DNA in it which caused a smear, however I am still highly confident that the results are accurate, that there is no Wolbachia in the common house cricket collected.  We had good controls for the remainder of the lanes in both gels.  We had a good DNA ladder, good arthropod positive and negative lanes even though there was a faint band in the Wolbachia gel.  There was a bright band for the positive DNA band.  There was nothing in the water lane.