Tubing prep station specifications, 1 precipitation of rna onto microcarriers – Bio-Rad Helios® Gene Gun System User Manual

Page 43

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Environmental

Operating

50 °F ( 10 °C) to 90 °F (32 °C) temp.

30–80 % humidity

Storage

32 °F (0 °C) to 140 °F (60 °C) temp.

10–90 % humidity

Tubing Prep Station Specifications

Physical

Dimensions

Approximately 33.5” x 4”

Weight

11.2 lbs

Construction

Aluminum and Acrylic

Electrical

Maximum Current

62 mA/125 mA peak

Voltage Input

220–240 Vac/100–120 Vac

Input Frequency

50/60 Hz

Functional

Input Pressure

30 psi N

2

Relief Pressure

30 psi ± 1.5 psi at regulator assembly

Regulator Adjustment

TDB psi limit maximum

Speed

30 RPM nominal

Tubing Fill

manual

Environmental

Operating

50 °F (10 °C) to 90 °F (32 °C) temp

30–80 % humidity

Storage

32 °F (0 °C) to 140 °F (60 °C) temp

10–90 % humidity

Section 10
Appendices

10.1 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 Section 5.2:

Purified RNA preparation (e.g. capped, polyadenylated, or oligo dT-selected mRNA).

5 M ammonium acetate

2-propanol

Experiments involving RNA require careful technique to prevent RNA degradation. In the stud-

ies published to date, it was not necessary to pre-treat the gold particles to prevent RNase contamination.

Procedure (sufficient for one length of Gold-Coat tubing):

1. In a 1.5 ml microfuge tube, weigh out 25 mg gold particles.

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