【Experimental Log】2016/02/17-18

17 Sample:CP210 (B. kawakamii), CP211 (B. kawakamii), CP212 (B. kawakamii), CP213 (B. kawakamii), CP214 (B. kawakamii), CP215 (B. kawakamii), CP216 (B. kawakamii), CP217 (B. kawakamii), CP218 (B. hayatana), CP219 (B. hayatana), CP220 (B. hayatana), CP221 (B. hayatana), CP222 (B. hayatana), CP223 (B. hayatana), CP230 (疑似B. pengii), CP231 (疑似B. pengii), CP232 (疑似B. pengii)

DNA extraction:CTab (未秤重,目視約一片正常大小葉子,加入液態氮磨碎。) 總體積用50 ug去溶。[CP230, CP231, CP232]

純化kit:Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB) [ALL]

DNA定量:Qbit

Qbit結果CP210 (B. kawakamii):100 ng/uL, CP211 (B. kawakamii):106 ng/uL, CP212 (B. kawakamii):100 ng/uL, CP213 (B. kawakamii):96.6 ng/uL, CP214 (B. kawakamii):99.2 ng/uL, CP215 (B. kawakamii):85.2 ng/uL, CP216 (B. kawakamii):79.8 ng/uL, CP217 (B. kawakamii):95.2 ng/uL, CP218 (B. hayatana):23.2 ng/uL, CP219 (B. hayatana):42.0 ng/uL, CP220 (B. hayatana):50.4 ng/uL, CP221 (B. hayatana):61.0 ng/uL, CP222 (B. hayatana):41.4 ng/uL, CP223 (B. hayatana):51.6 ng/uL, CP230-1 (疑似B. pengii):70 (68.8)ng/uL, CP230-2 (疑似B. pengii):80.2 (79.8) ng/uL, CP231-1 (疑似B. pengii):104 ng/uL, CP231-2 (疑似B. pengii):96.8 ng/uL, CP232-1 (疑似B. pengii):110 (108) ng/uL, CP232-2 (疑似B. pengii):91 ng/uL

結論:大致上狀況都還不錯,都在預期內。

CP230, CP231, CP232是新來的。登山社學長楊斯顯從台灣東南部美奈田山帶回來的。正好是我很缺少的取樣點 (連油脂都沒去過耶,他只聽過那裏有小檗,但不知道是誰)。拿到的後有很多份,但枯得差不多了,但其中有花有果的狀況竟然算好。清楚看到花的最外一輪花萼為尖形,果為圓,加上地理位置,學長推斷他為南台灣小檗 (B. pengii)。但我對南台灣小檗實在不熟悉,無法一時間認出來。利馬用液態氮磨碎抽DNA。
CP210~217的Sample,阿倫在20151207幫我在大雪山採,以及老師在20151208在塔塔加幫我採的台灣小檗 (B. kawakamii)。那時候剛拿到就已經抽好了,一直沒有過colum和測濃度,就剛好用這組新的純化Kit,也剛好有Qbit,就一起做完。
CP218~223的Sample,是在20151214和家人一起去太平山採回來的。一樣當時已經抽好,到屆時才純化+測濃度。
昨天(20160217)翊萱幫我從向陽帶小檗回來。和學長討論後感覺應該是南台灣小檗 (B. pengii)。我沒有辨識成功。對他的多樣化似乎還沒有抓到精隨。明天(20160219)買到液態氮再一口氣做好DNA吧。
冰箱裡還有幾支落單的DNA,找時間也過純化kit+測濃度後再收進盒裡。還有陽台上的活體小檗,也大概要收DNA了。這些事情希望能在下禮拜二,油脂學長回來之前完成。(還要討論出國採集一事,忙碌的一學期要開始了)

加油。

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