【Experimental Log】2016/02/16

11 Sample:CP162 (B. tarokoensis), CP163 (B. tarokoensis), CP164 (B. tarokoensis), CP164n (B. tarokoensis), CP165 (B. tarokoensis), CP166 (B. tarokoensis), CP167 (B. tarokoensis), CP168 (B. tarokoensis) [8/10抽的藍字蓋那批 (有兩組,另一組為黑字蓋)], CP185 (B. tarokoensis), CP186 (B. tarokoensis), CP187 (B. tarokoensis)

純化kit:Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB) (之前的Monarch DNA Gel extraction kit,是用在切膠上,這是直接拿來清DNA使用。)

DNA定量:Qbit


 

Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB)

原始:CP162 (B. tarokoensis), CP163 (B. tarokoensis), CP164 (B. tarokoensis), CP164n (B. tarokoensis), CP165 (B. tarokoensis), CP166 (B. tarokoensis), CP167 (B. tarokoensis), CP168 (B. tarokoensis),用nanodrop測的時候,品質就已經很糟了,依然留有視覺髒色,也沒有濃度 (詳見:https://champ1115.wordpress.com/2015/08/14/%E3%80%90experimental-log%E3%80%91201587-9/)。

過程:全使用Cleanup Kit,依造protocol。

結果:全無。原因可能是本來在過Colum就把DNA通通洗光了。


Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB)

原始:CP185 (B. tarokoensis):Too high, CP186 (B. tarokoensis):4.78 (5.04) ng/uL, CP187 (B. tarokoensis):87.6 ng/uL

過程:全使用Cleanup Kit,微調protocol,只洗一次。

結果:CP185 (B. tarokoensis):84.6 ng/uL, CP186 (B. tarokoensis):0.18 ng/uL, CP187 (B. tarokoensis):64 ng/uL

結論:

經前後比較測試可發現,最多會掉約20 ng/uL。但也有可能為Qbit測濃度的誤差。但重點是:1.原來濃度足夠使用上機的,Cleanup完後,依然可以。2.髒污或視覺髒色消失。

足矣。

廣告

發表迴響

在下方填入你的資料或按右方圖示以社群網站登入:

WordPress.com Logo

您的留言將使用 WordPress.com 帳號。 登出 / 變更 )

Twitter picture

您的留言將使用 Twitter 帳號。 登出 / 變更 )

Facebook照片

您的留言將使用 Facebook 帳號。 登出 / 變更 )

Google+ photo

您的留言將使用 Google+ 帳號。 登出 / 變更 )

連結到 %s