BIO RAD Helios Gene Gun System, Instruction Manual

Page: 54 / 60

Section 10 Appendices
50 | Helios Gene Gun System
Quantification of DNA in Cartridges
This procedure can be used for estimating the amount of DNA in cartridges when 0.5 µg or more of DNA
has been loaded per tube.
Materials
■
10 mM Tris, pH 8.0, 1 mM EDTA
■
1,000 µ micropipettor and tips
■
Ultrasonic cleaner
■
UV spectrophotometer
■
Vortexer
■
Quartz cuvettes
■
Microfuge
Procedure
1. Place five 0.5" cartridges in a microcentrifuge tube and add 500 µl of TE.
2. Sonicate and/or vortex to dislodge gold and solubilize the DNA. Spin for 5 min in a microfuge to
pellet gold.
3. On a spectrophotometer, determine the A
260
by reading against a TE blank. If 100% of the DNA were
precipitated onto the microcarriers, the A
260
readings should be as follows:
Plasmid/Cartridge, µg A
260
0.5 0.1
1.0 0.2
2.0 0.4
4. DNA sample may also be analyzed by restriction enzyme digestion and analysis by agarose
gel electrophoresis.