Harmer Lab:Protocols:Direct PCR: Difference between revisions
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Modified from [http://www.amjbot.org/cgi/content/abstract/97/7/e65| Bellstedt et al. 2010] by Matt | Modified from [http://www.amjbot.org/cgi/content/abstract/97/7/e65| Bellstedt et al. 2010] by Matt | ||
#Add a single ball bearing to each 1.2 ml tube containing tissue ( | #Add a single ball bearing to each 1.2 ml tube containing tissue (1 cotyledon sized piece works, up to 1 small leaf normally) | ||
#Add 350μL grinding buffer | #Add 350μL grinding buffer | ||
#Disrupt tissue by shaking with paint shaker for 1 min. Repeat if clumps of tissue remain | #Disrupt tissue by shaking with paint shaker for 1 min. Repeat if clumps of tissue remain | ||
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#2M NaCl, 2.5mL | #2M NaCl, 2.5mL | ||
#0.5M EDTA, pH8 200μL | #0.5M EDTA, pH8 200μL | ||
# 500μL Triton X-100 | |||
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Latest revision as of 12:58, 3 November 2011
Room 2123 |
96-Well Format DNA Extraction for Direct PCRModified from Bellstedt et al. 2010 by Matt
100mL Grinding BufferTo prepare 100 mL of grinding buffer dissolve all components except Tween 20 in double-distilled sterile water. Stir at low speed to avoid excessive foaming. Once all components have dissolved, adjust pH to 9.6 using NaOH solution. Then add Tween 20. Autoclave and store at 4°C.
100mL GES Buffer
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