Measurement

IDT order primer concentrations when arrived

Diluted each primer to 100 μM by adding concentration x 10 • Eg. 27.3 nmol added with 273 μL of water for a final concentration of 100 μM

Before PCR; EHMT2 was in an agar stab

After Gel Extraction

Testing EHMT2 colonies (after plasmid extraction)

Testing New vs Old EHMT2 colonies

Testing EHMT2 new PCR product colonies (N) vs old colonies (O)
Key: EHMT2 N1 indicates the 1st new colony for EHMT2

PCR Products

Gibson Assembly, cut with restriction enzymes

First PCR with the GFP 3.3, Puro, and EHMT2 as an agar stab
The following PCRs follow the same measurements as this one; varying depending only on the DNA concentration

Mass of each piece of gel after electrophoresis

First Gibson Assembly run:

Gibson Assembly for New vs Old EHMT2

4 fragment Assembly, recommended by NEB

Conducted 2 double digests, following the same format

Before plating the cells into the wells, we measured the confluency and amount of cells per mL

Plated 6 plates of cells in this measurement; 3 of the plates were cells transfected with the plasmid, and 3 of the plates were cells not transfected with the plasmid.

The bottom right corner 3 wells were left blank as a control. We transfected siRNA into all 6 plates.

Light blue - transfected 3 days
Dark blue - transfected 4 days
Orange - cells without the plasmid, transfected 3 days

Same measurements for all wells containing samples
(components & measurement here are in per well for a standard qPCR)