PCR from Plant Tissue

1.protocol

(1)collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH

(2)put in boiling H₂Ofor 30 sec (optimum may need to be determined for each type of tissue, for floral tissue 30 sec is fine)

(3)neutralize by adding 40 ml 0.25 N HCl and 20 ml 0.5 M Tris-HCl pH 8.0, 0.25% (v/v) Igepal CA-630

(4)boil for another 2 min

(5)use directly for PCR or store at 4°C for up to several weeks

(6)if stored at 4°C boil again for 2 min before the PCR

2.Remarks

We don't have too much experience with this method. Mostly it seems to work though.However, I don't think it is safe enough for critical purposes (e.g., mapping).

3.Materials

Reagent	Supplier	Cat.-#
Igepal CA-630	Sigma	I-3021

4.Reference

Klimyuk et.al., 1993, Plant J. 3:493-494