Bio-Rad Sub-Cell® Model 192 Cell - Manual (Page 18)

1x Tris-Acetate-EDTA (TAE) — 40 mM Tris (pH 7.6), 20 mM Acetic Acid, and 1 mM EDTA.
50x Stock (1 liter):
dissolve in 600 ml distilled water:
Tris Base (FW = 121)
242.0 g
Glacial acetic acid 57.1 ml
0.5 M EDTA (pH 8.0) 100.0 ml
Fill to a final volume of one liter with distilled water.
1x Tris-Boric Acid-EDTA (TBE) — 89 mM Tris (pH 7.6), 89 mM Boric Acid, 2 mM EDTA
10x Stock (1 liter):
dissolve in 600 ml distilled water:
Tris Base (FW = 121)
108 g
Boric Acid (FW = 61.8) 55 g
0.5 M EDTA (pH 8.0) 40 ml
Fill to a final volume of one liter with distilled water.
1x MOPS Buffer (RNA Gels) — 0.02 M MOPS [3-(N-morpholino)-propanesulfonic acid]
(pH 7.0), 8 mM Sodium Acetate, 1 mM EDTA (pH 8.0)
5x Stock (1 liter):
dissolve in 600 ml DEPC-treated distilled water:
MOPS
20.6 g
3 M Sodium Acetate (DEPC treated) pH 7.4 13.3 ml
0.5 M EDTA (DEPC-treated) pH 8.0 10.0 ml
Fill to a final volume of one liter with DEPC-treated distilled water.
Caution: DEPC is a suspected carcinogen. Always wear gloves, eye glasses and a
laboratory coat. Use caution when handling DEPC containing solutions. Consult the
DEPC MSDS (Material Safety Data Sheet) for more information.
3.2 DNA and RNA Gel Preparation
DNA Agarose Gels
(See Section 2.1)
RNA Agarose Formaldehyde Gels 1-2
For 100 ml of a 1% agarose formaldehyde gel prepare as follows:
1.6% melted agarose 62 ml
5x MOPS Electrophoresis Buffer (1x final concentration) 20 ml
12.3 M (37.5%) Formaldehyde (2.2 M final concentration) 18 ml
Caution: Formaldehyde solutions and formaldehyde vapors are toxic. When handling
solutions or gels that contain formaldehyde use a chemical hood. Always wear gloves, eye
glasses and a laboratory coat while using formaldehyde. See the formaldehyde MSDS
for more safety information.
3.3 RNA Sample Preparation 1-2
Prior to loading RNA onto an agarose formaldehyde gel prepare each RNA sample as
follows:
3.0 µl RNA in DEPC-treated water
5.0 µl 5x MOPS Buffer (final concentration 1.67x)
4.5 µl 12.3 M Formaldehyde (final concentration 3.7 M)
12.5 µl Formamide (final concentration 50% v/v)
Caution: Formamide is a teratogen. Always wear gloves, eye glasses and a laboratory
coat. Use caution when handling formamide. Consult the formamide MSDS for more
information.
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