Bio-Rad Model 785 Vacuum Blotter - Manual (Page 10)

2. There are three transfer procedures listed: the Standard Transfer Procedure, the
Rapid Transfer Procedure, and the RNA Transfer Procedure. The Standard Transfer
Procedure is for detection of single copy gene in genomic DNA. The Rapid Transfer
Procedure is for fast identification of DNA inserts from various cloned vectors.
Standard Procedure:
1. Depurinate the gel in 0.25 N HCl for 15 minutes in a tray. Cover the gel with 0.25 N
HCl and shake gently.
2. Remove the 0.25 N HCl solution. Rinse the gel twice with deionized distilled water.
3. Denature the gel in 0.5 N NaOH for 30 minutes. Cover the gel with 0.5 N NaOH and
shake gently.
4. Transfer the gel in 10x SSC for 90 minutes at 5 inches Hg. Follow the instructions in
Section 3.4 for the transfer procedures.
Rapid Transfer Procedure:
1. Depurinate the gel in 0.25 N HCl for 15 minutes in a tray. Cover the gel with 0.25 N
HCl and shake gently.
2. Remove the 0.25 N HCl solution. Rinse the gel twice with deionized distilled water.
3. Immediately, transfer in 0.5 N NaOH, 0.6 N NaCl for 90 minutes at 5 inches Hg.
Follow the instructions in Section 3.4 for the transfer procedures.
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Fig. 3.2. Exploded view of vacuum transfer set-up.
Sealing frame
Agarose gel
Window gasket
Nylon membrane
Filter paper
Porous vacuum plate