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Originally posted by wangleisdnu at 2010-12-04 00:34:11:
normally I follow this protocol
E. coli Colony PCR
1. Pick colony with toothpick and swirl into 25 ¦Ìl of TE (or dH2O) in microfuge tube.
2. Heat in boiling water bath for 2 minutes (can be use ...

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5Â¥2010-12-04 16:26:28
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dhd997(½ð±Ò+3):good 2010-12-04 08:22:16
normally I follow this protocol
E. coli Colony PCR
1. Pick colony with toothpick and swirl into 25 ¦Ìl of TE (or dH2O) in microfuge tube.
2. Heat in boiling water bath for 2 minutes (can be used directly for PCR).
3. Spin for 2 minutes at top speed in microfuge.
4. Transfer 20 ¦Ìl of supernatant to new microfuge tube.
5. Use 1-2 ¦Ìl as template in 25 ¦Ìl PCR reaction.

in my hands it works well


maybe your culture concentration is too high, which affects the PCR reaction
ÓÐÓÂÆøÈ¥¸Ä±äÄã¿ÉÒԸıäµÄ,ÓжÈÁ¿È¥ÈÝÈÌÄã²»ÄܸıäµÄ,ÓÐÖÇ»ÛÈ¥Çø·ÖÉÏÃæµÄÁ½ÖÖÇé¿öhttp://bbs.bioon.net/bbs/?66210
2Â¥2010-12-04 00:34:11
ÒÑÔÄ   »Ø¸´´ËÂ¥   ¹Ø×¢TA ¸øTA·¢ÏûÏ¢ ËÍTAºì»¨ TAµÄ»ØÌû

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