| PCR from Plant Tissue | | 点击： 作者： 来源： 时间： 2006-11-06 本站论坛 |
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PCR from Tissue
Reference: Klimyuk et.al., 1993, Plant J. 3:493-494 Last updated: 1/27/00 By: Kay Schneitz
- collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH
- put in boiling H2O for 30 sec (optimum may need to be determined for each type of tissue, for floral tissue 30 sec is fine)
- 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
- boil for another 2 min
- use directly for PCR or store at 4°C for up to several weeks
- if stored at 4°C boil again for 2 min before the PCR
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).
Materials:
Reagent Supplier Cat.-#
Igepal CA-630 Sigma I-3021
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