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DNA Cleanup & Plasmid Purification critical

No DNA Recovered from Gel Extraction or PCR Cleanup

Symptom
No DNA is recovered after gel extraction or PCR cleanup procedures. Elution yields no measurable nucleic acid.
Common Causes
  1. 1 Ethanol not added to DNA Wash Buffer before use (Monarch DNA Gel Extraction Kit #T1020 or PCR Cleanup Kit #T1030)
  2. 2 Buffers not reconstituted correctly before starting procedure
  3. 3 Reagents added in incorrect order disrupting binding or elution
Solutions
  1. 1 Verify proper amount of ethanol was added to Monarch DNA Wash Buffer per kit protocol
  2. 2 Check that all buffers are reconstituted according to kit instructions before use
  3. 3 Follow reagent addition order precisely: sample preparation → binding → washing → elution
Related Video (2)
New England Biolabs ★ 90
Monarch PCR & DNA Cleanup Kit Protocol
"Directly demonstrates the Monarch PCR & DNA Cleanup Kit protocol, including correct buffer preparation to avoid the ethanol addition failure."
QIAGEN ★ 65
Protocol: Plasmid DNA Purification using the QIAprep Spin Miniprep Kit and a Vacuum Manifold
"Shows a similar bind-wash-elute plasmid purification workflow, helping users understand the general process and reagent steps where ethanol is required."
Source: neb.com ↗
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