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Trouble-shooting: Low yield

ability and work equally well in PCR, sequencing, and labeling applications. Lithium salts are more soluble in ethanol than sodium salts. Thus, removal of lithium salts by ethanol precipitation is more efficient than removal of sodium salts. Using lithium salt nucleotide preparations reduce salt-induced artifacts and increase the legibility of sequencing gels. Back to Top Nucleotide concentration is too high or unbalanced

The standard concentration is 20-200 M of each nucleotide

Suggestions:

  1. Check the concentration of stock solutions of all nucleotides
  2. Double check the final concentrations of all nucleotides
Back to Top Primer concentration is too low

Suggestion:

Adjust the primer concentration (0.1 - 1.0 M of each primer is optimal)

Back to Top

Machine based error Suggestions:
  • Calibrate the heating block and confirm actual block temperature
  • Run a diagnostic program for the particular machine - contact the machine manufacturer for specifics
Back to Top
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