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DIG Southern Blot Solutions

0.25M HCl (1L)

• 20.66mL HCl
• 979.34mL dH₂O
• *add acid to water*

1M ammonium acetate, 0.02M NaOH (1L)

• 77.08g ammonium acetate
• 0.8g NaOH
• Fill with dH₂O

0.5X SSC, 0.1% SDS (1L)

• 25mL 20X SSC
• 10mL 10% SDS
• 965mL dH₂O

10X Blocking Buffer Þ 1X (30mL)

• 3mL Blocking Buffer stock
• 27mL Maleic Acid Buffer

Wash Buffer 3 (30mL)

• 30mL Maleic Acid Buffer
• 90µ l Tween 20 (cut off pipet tip)

Detection Buffer (1L)

• 100mL 1M Tris pH 9.5
• 20mL 5M NaCl
• 880mL dH₂O
• *Autoclave*

1.5M NaCl, 0.5M NaOH (1L)

• 87.66g NaCl
• 20g NaOH
• Fill with dH₂O

2X SSC, 0.1% SDS (1L)

• 100mL 20X SSC
• 10mL 10% SDS
• 890mL dH₂O

Maleic Acid Buffer (1L)

• 11.61g Maleic Acid
• 8.766g NaCl
• Fill with dH₂O
• *check pH 7.5 with NaOH pellets (~7g)
• *Autoclave*

Diluting Ab 1:10,000 (30mL)

• 3mL Blocking Buffer stock
• 27mL Maleic Acid Buffer
• 3µ l Ab