DyNAzymeTM EXT DNA Polymerase Blunt vs' TAcloning

1
DyNAzymeTM EXT DNA Polymerase Blunt vs.
TA-cloning

• Purpose of the test
• To study what kind of PCR products DyNAzyme EXT
  produces (blunt end/A-overhang) in order to find
  out how well it can be used for different cloning
  applications.
• Principle of the cloning test
• Different type of primers were used for
  amplification in order to find out what kind of
  effect the different primer sequences have to
  the end product.

2
Blunt vs. TA-cloning
3
Conclusions

• DyNAzyme EXT works well both in blunt-end and
  TA-cloning.
• Both blunt and "overhanged" PCR products were
  produced during PCR with 9 different primers.
• Some general rules can be taken out from these 9
  examples.
• If the last nucleotide of the PCR product is an
  adenine (the PCR primer 5 end starts with a
  thymidine) then the non-template addition of A is
  highly unlikely and blunt cloning can be
  recommended.
• If the PCR primer starts with an A (PCR product
  3'-end is T) then DyNAzyme EXT prefers A-addition
  and TA-cloning can be recommended.
• Same kind of results are described in
  BioTechniques by V.L. Magnuson et al (vol 21
  700-709, No4,1996).