BIO RAD Helios Gene Gun System, Инструкция по эксплуатации

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Section 8 Troubleshooting
Helios Gene Gun System | 39
Parameters for In Vivo Delivery
The following are suggested starting conditions for optimizing DNA delivery to the skin of various species.
Each laboratory should determine the optimum parameters for their particular application (see Overview in
Section 7 and Table 5). Bombardment conditions are those used in references cited in Section 10, using the
Accell Gene Gun. Use this information as a starting point when optimizing bombardment conditions with the
Helios Gene Gun. Lower helium pressures should be tested early in the optimization process.
Table 5. Examples of parameters for delivery into skin of various species
Species Site MLQ, mg/target DLR, µg/mg PVP, mg/ml psi
Dog
9
Dorsum 0.5 1–2 0.1 300
Monkey
7, 8, 9
Abdomen 0.25 2 0–0.05 300–400
Mouse
17, 19
Abdomen* 0.25–0.5 1–2.5 0–0.1 300–500
Pig
7, 8, 10
Inner thigh (clipping not
necessary)
0.25–0.5 0.5–2.5 0.05 500
Rabbit
27
Back 0.5 1–2 None 350–400
* Do not use a depilatory agent to remove the fur (Section 6.3, step 3)
Section 8
Troubleshooting
DNA/Microcarrier Preparation
Problem Microcarriers agglomerate after coating with DNA.
Possible solutions Lower DNA loading rate.
Cartridge Preparation
Problem Gold does not spread evenly in the GoldCoat Tubing
(rings, clumps, uncoated sections, streaks).
Possible solutions Eliminate any potential sources of water in the tube and in the final DNA/
microcarrier preparation.
Rewash remaining microcarrier prep 2–3 times using a fresh bottle of
100% ethanol.
Flush GoldCoat Tubing with nitrogen gas for 15 min prior to loading with
DNA/gold microcarrier solution.
If using PVP, make a new stock of PVP in ethanol using a fresh bottle of alcohol.
Resuspend gold/DNA pellet prior to first ethanol wash.
After drawing off the ethanol, quickly turn the GoldCoat Tubing with your
fingers prior to rotating.