Bio-Rad Zeta-Probe Membranes User Manual

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Section 1
Introduction

Zeta-Probe blotting membranes are nylon membranes which have
unique binding and handling properties that make them ideally suited
for nucleic acid, and some protein, blotting applications.

Zeta-Probe membranes possess a high tensile strength. They won't
shrink, tear, or become brittle during transfer, baking, hybridization, or
reprobing. Zeta-Probe membranes are heat-resistant, nonflammable,
and autoclavable. Zeta-Probe membranes are naturally hydrophilic
with no added wetting agents. These membranes are resistant to a
wide variety of chemicals, including 100% formamide, 2 M NaOH, 4
M HCl, acetone, most alcohols, DMSO, DMF, and chlorinated aliphat-
ic hydrocarbons. The nominal porosity of Zeta-Probe membranes is
0.45 µm. When stored at 23–25°C, Zeta-Probe membranes are
stable for at least 1 year.

When handling Zeta-Probe membranes, always wear gloves or use
forceps. After blotting, do not allow wet membranes to come in con-
tact with each other. Contact may result in the transfer of blotted
nucleic acids or proteins from one membrane to the other.

Stock buffers are listed in the appendix. It is suggested that you read
the entire protocol before proceeding.

Section 2
Nucleic Acid Blotting Protocols

Several nucleic acid blotting methods are presented in this section.
Capillary blotting (Sections 2.1through 2.3) is generally used with
agarose gels, and electrophoretic transfer (Section 2.4) is used with
polyacrylamide gels. Dot blotting (Sections 2.5 and 2.6) is used for

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