Bob Marcil r_marcil at INVITROGEN.COM
Tue Nov 25 01:12:53 EST 1997

                                                                      11:14 PM

Although the idiots at Invitrogen won't allow me to incorporate these methods
into a new kit, I would like to endorse two techniques that can greatly
enhance the success of PCR cloning, especially for long PCR products. Yields
of mix polymerase PCR (Taq + a proofreading enzyme ~20:1 (Barnes, W. PNAS 91,
1994 p2216-2220)) can be increased by using 1.3 M anhydrous betaine (ref:
Hengen, P. N., TIBS 22 June, 1997 p225-226). Improved cloning efficiencies by
avoiding UV exposure can be achieved by using crystal violet during agarose
gel electrophoresis purification (ref: Rand, K.N., Elsevier Trends Journals
Technical Tips Online http:/tto.trends.com/cgi-bin) These techniques really
work. Enjoy!  --Bob

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