Tubing Prep Station Specifications; Appendices; Precipitation Of Rna Onto Microcarriers - BIO RAD Helios Gene Gun System Instruction Manual

Tubing Prep Station Specifications

Specifications
Physical
Dimensions Approximately 33.5 x 4"
Weight 11.2 lb
Construction Aluminum and acrylic
Electrical
Maximum current 62 mA/125 mA peak
Voltage input 220–240 V AC/100–120 V AC
Input frequency 50/60 Hz
Functional
Input pressure 30 psi N
Relief pressure 30 ± 1.5 psi at regulator assembly
Regulator adjustment TDB psi limit maximum
Speed 30 RPM nominal
Tubing fill manual
Environmental
Operating 50–90°F (10–32°C) temp
30–80 % humidity
Storage 32–140°F (0–60°C) temp
10–90 % humidity
Section 10

Appendices

Precipitation of RNA onto microcarriers

Primary RNA transcripts and mRNA may be effectively delivered in vivo and in vitro using the Helios
Gene Gun System (Qiu et al., 1996). The procedure is similar to that for DNA, but the precipitation step is
performed with ammonium acetate and 2-propanol.
Materials
In addition to those identified in Preparation of System Components Prior to Bombardment, Section 5:
Purified RNA preparation (for example, capped, polyadenylated, or oligo dT-selected mRNA).
■
5 M ammonium acetate
■
2-propanol
■
Experiments involving RNA require careful technique to prevent RNA degradation.
Procedure (sufficient for one length of GoldCoat Tubing):
1. In a 1.5 ml microcentrifuge tube weigh out 25 mg gold particles.
2. To measured gold add aqueous solution of mRNA preparation. The ratios of RNA to gold used are
similar to those used for DNA (1–15 µg RNA/mg particles).
3. Add 1/10 volume of 5 M ammonium acetate and 2 volumes of 2-propanol, mix by vortexing. Place the
microfuge tube at -20°C for 1 hour.
4. Proceed with Section 5.2, steps 9–12.
2
Section 10 Appendices
Helios Gene Gun System | 43