Klenow Fill-In Kit

Choose the Klenow Fill-In Kit to perform both partial fill-in reactions to complete fill-in of a 5 overhang to generate blunt ends, and allow direct ligation. Or, use the kit to radioactively end label a 5 overhang with [alpha-32P]dNTP. The Klenow Fill-in Kit contains Klenow fragment (125 U), dNTPs (separate 10 mM stocks), Klenow reaction buffer, predigested DNA, which can be used as a control, and sufficient material for 25 reactions. Perform a complete fill-in of a 5 overhang to generate blunt ends, to allow direct ligation Radioactively end label a 5 overhang with [alpha-32P]dNTP Includes predigested DNA that can be used as a control The sample DNA should be extracted with phenol–chloroform and ethanol-precipitated before performing a fill-in reaction. Add the Klenow polymerase to the reaction mixture last—after the DNA, fill-in buffer and dNTPs—and do not let the reaction mixture sit on ice for an extended period of time.