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PCR from Plant Tissue

2007-7-8 22:58:54 信息来源:来源网络 
  •   PCR from Plant Tissue
http://www.bioon.com 生 物 谷 网 站PCR from Tissue



Reference:  Klimyuk et.al., 1993, Plant J. 3:493-494 Last updated: 1/27/00 By: Kay Schneitz

     



 
  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 H2O for 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

 

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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