Bio-Rad Trans-Blot® Cell - Manual (Page 12)

Table 3.2. DNA and RNA
Standard Field Standard Field High Intensity Field
Overnight 8 cm electrode distance 4 cm electrode distance
❄
COOLING REQUIRED
TAE or TBE TAE TBE TAE TBE
Trans-Blot cell with 30 V, 0.2 A 60 V, 0.6 A 80 V, 0.55 A 80–100 V 100–130 V
wire electrodes 0.8–1.4 A 0.7–1.1 A
Overnight 5 hours approximately 30 minutes–4 hours ❄
Trans-Blot cell with TAE or TBE TAE TBE TAE TBE
plate electrodes 15 V, 0.2 A 20–35 V, 75–100 V 15–25 V 50–90 V
0.8–1.5 A 0.8–1.1 A 0.8–1.4 A 0.7–1.1 A
Overnight 1–5 hours ❄ 30 minutes–1 hour ❄
1x TAE: 40 mM Tris-Acetate, 1.0 mM EDTA
1x TBE: 90 mM Tris-Borate, 2.0 mM EDTA
Table 3.3. Native Gels
Standard Field Standard Field High Intensity Field
Overnight 8 cm electrode distance 4 cm electrode distance
❄
COOLING REQUIRED
Trans-Blot cell with 30 V, 0.1 A 60 V, 0.21 A 100–200 V
wire electrodes 0.3–0.8 A
Overnight 5 hours approximately 30 minutes–4 hours ❄
Trans-Blot cell with 10 V 100–150 V 50–100 V
plate electrodes 0.1 A 1–1.5 A 0.7–1.5 A
Overnight 1–5 hours ❄ 30 minutes–1 hour ❄
Buffer: 25 mM Tris, pH 8.3,192 mM glycine. No methanol.
Table 3.4. Isoelectric Focusing, Native Gels, Basic Proteins, Acid Urea Gels
Standard Field Standard Field High Intensity Field
Overnight 8 cm electrode distance 4 cm electrode distance
❄
COOLING REQUIRED
Trans-Blot cell with 30 V, 0.2 A 70 V, 0.5 A 100–150 V, 0.55–0.85 A
wire electrodes Overnight 5 hours approximately 30 minutes–4 hours ❄
Trans-Blot cell with 15 V, 0.2 A 40–70 V, 0.6–1.0 A 30–-60 V, 0.6–1.0 A
plate electrodes Overnight 1–5 hours
❄ 30 minutes–1 hour ❄
Buffer: 0.7% Acetic Acid
3.2 Notes on Electrophoretic Transfer Conditions
These variables will change total resistance and thus the current readings:
• Alterations in buffer make-up, i.e., addition of SDS, or changes in ion concentration
due to addition of acid or base to adjust the pH of the buffers.
• Gel pH, ionic strength, and percentage of acrylamide, especially if the gel has not
been properly equilibrated.
• Number of gels; current increases slightly as the number of gels increases.
• Volume of buffer; current increases when volume increases.
• Platinum mass; current increases when mass increases.
• Transfer temperature; current increases when temperature increases.
• Time in transfer at which reading was taken; current normally increases as the buffer-
ing capacity diminishes with progress of the run.
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