Q9.  IS IT POSSIBLE TO CARRY OUT LA PCR DIRECTLY ON

                PHAGE PARTICLES?

It is possible to generate DNA fragments in lengths of 8 kbp or longer when using 10 to

10 PFU particles prepared with heat treatment at 99 for 10 minutes as templates.

LA PCR amplification using phage particles prepared with heat treatment as

a template

When amplified with the LA PCR Kit Ver. 2.1, fragments of 8 kbp or longer can be

generated, while the conventional PCR Amplification Kit using a normal Taq can

generate fragments up to 6 kbp in length with poor yield.

Reactions were conducted under optimal conditions following the directions provided

with the kits (LA PCR Kit Ver. 2.1 and PCR Amplification Kit) for the amplification of
phage templates. phage particles were prepared by treating 10⁷ PFU of a phage

suspension in SM* buffer with heat at 99 for 10 minutes.

NaCl                      

5.8 g

MgSO₄ 7H₂O

2 g

1 M Tris HCl (pH7.5) 

50 ml

2% gelatin solution      

5 ml

Applied volume:

6 

1% Agarose gelelectrophoresis