【Experimental Log】2015/7/15-16

6 Sample:886 (B. kawakamii), CP98 (B. hayatana), CP103 (B. kawakamii), CP124 (B. kawakamii), CP137 (B. schaaliae), CP141 (B. chingshuiensis)

DNA extraction:CTab (未秤重,目視約兩片正常大小葉子,揉碎直接用鋼珠打碎。) 在65度時放入搖晃機中,約莫一小時,此次只用震盪機 (50 1/s*3 mins)*3 (一顆大、兩顆中鋼珠),開始前要將葉子先撥碎。總體積用50 (-1) ug去溶:886 (-2)。皆有過Colum。

NanoDrop (曲老師實驗室)886 (B. kawakamii):389.7 ug/mL, 1.89 260/280, 2.18 260/230, CP98 (B. hayatana):66.6 ug/mL, 1.89 260/280, 2.52 260/230, CP103 (B. kawakamii):75.4 ug/mL, 1.88 260/280, 2.09 260/230, CP124 (B. kawakamii):177.8 ug/mL, 1.83 260/280, 2.17 260/230, CP137 (B. schaaliae):73.4 ug/mL, 1.87 260/280, 2.36 260/230, CP141 (B. chingshuiensis):41.7 ug/mL, 1.8 260/280, 2.06 260/230

Note:告一個小小段落。最近有點為低潮,卻沒有來由。暑假很不像暑假。老妹考試加油吧。

【Experimental Log】2015/7/13-14

8 Sample:CP82 (B. aristatoserrulata), CP88 (B. aristatoserrulata), CP100 (B. hayatana), CP101 (B. kawakamii), CP123 (B. nantoensis), CP132 (B. schaaliae), CP139 (B. chingshuiensis), CP142 (B. chingshuiensis)

DNA extraction:CTab (未秤重,目視約兩片正常大小葉子,揉碎直接用鋼珠打碎。) 在65度時放入搖晃機中,約莫一小時,此次只用震盪機 (50 1/s*3 mins)*3 (一顆大、兩顆中鋼珠),開始前要將葉子先撥碎。總體積用50 (-1) ug去溶:CP82 (-2)、CP139 (-3)、CP93 (-2,包括上次已 -3)。皆有過Colum。

NanoDrop (曲老師實驗室)CP82 (B. aristatoserrulata):134.9 (134.1) ug/mL, 1.85 (1.85) 260/280, 2.06 (2.06) 260/230, CP88 (B. aristatoserrulata):86.9 ug/mL, 1.85 260/280, 2.19 260/230, CP100 (B. hayatana):48.3 ug/mL, 1.87 260/280, 2.01 260/230, CP101 (B. kawakamii):54.1 ug/mL, 1.80 260/280, 1.81 260/230, CP123 (B. nantoensis):99.8 ug/mL, 1.84 260/280, 1.89 260/230, CP132 (B. schaaliae):99.2 ug/mL, 1.82 260/280, 2.09 260/230, CP139 (B. chingshuiensis):94.9 (75.4) ug/mL, 1.73 (1.87) 260/280, 1.51 (2.11) 260/230, CP142 (B. chingshuiensis):59.5 ug/mL, 1.79 260/280, 1.57 260/230。[重測CP93 (B. ravenii):91.1 (87.8) ug/mL, 1.81 (1.85) 260/280, 1.87 (1.87) 260/230。]

Note:這次全部是抽自己的編號。把一些有缺的種類,盡量找出來,抽掉。這次長葉小檗 (B. aristatoserrulata) 抽了兩支,一支是嵐山鐵道的,一支是ㄓㄐ學長從小鬼湖帶回來的,保存都還算OK。至於南投小檗 (B. nantoensis),這一支是自己從合歡山小奇萊帶回來的,都小片小片,結果竟還不錯,這和保存方法可能有關係 (下面會談)。清水山小檗 (B. chingshuiensis),好可怕,活體活的狀況有點糟,剩下的sample也普普通通,就把剩下的抽完,幸好還有抽出東西,尤其CP139上上次有抽過,所以sample只剩下一小片,但上次狀況很糟,這次還算OK。至於,CP93為什麼重測,原因很北七,上次做完實驗沒有冰到他,就讓它裸在室溫,看到我就傻眼。幸好還有東西出來,希望不要斷裂得太誇張。CP139測濃度用量為何用到-3,因為沒有混勻就測導致第一次測濃度竟然是0!之後就有,但是數值變化很大,但有已經花掉3了,不太敢再測下去。

[20150714和ㄓㄐ臉書討論] Q:倫太文山採的那批高山和長葉 是大概幾月 然後你是怎麼保存得 是一踩就直接丟矽膠 還是 採了最後才丟矽膠?(這是我覺得會影響sample保存的關鍵) A:四月中 沒有當場丟矽膠,在採集當晚才丟到矽膠裡。(我覺得這是影響保存的重點)  而且矽膠量少,因為我不想背,然後回來後才補加。[ㄓㄐ的想法]:沒有耶,因為我覺得小檗葉片的乾燥包存看似沒有pattern,那次梅雨季環境濕氣重,所以基本上還以為包存會不好 (但那批其實保存的很好),DNA很容易受水分破壞,小檗是革質葉,我猜矽膠乾燥的效率很動態,有些可能還有加強的厚壁組織,會防止水分喪失。[我的想法]:我現在有一個經驗,是上次去清水山的時候很明顯的。我在路上看到第一棵花蓮小檗,我馬上就進矽膠,但之後花蓮就越來越多,其他人要前進就也沒辦法一直等我,所以我都先採枝條,最後晚上再放,結果直接放的那份全黑。其餘得到現在保存還很好。像上次去眠月,都是直接放矽膠,情況都很糟。因為倫太文山那批高山小檗,很讓我意外。因為那是很難保存的,卻都很好,抽出來濃度都還很高,而且汙染很少。又尤其時間又過了一段時間了。→似乎可以再討論看看,想探究原因到底為何。(像這次抽的sample裡的南投小檗也是,那時候在小奇萊大雨,採枝條回住的地方才放矽膠,(比起之前抽南投的經驗) 情況算是不錯。)

【Experimental Log】2015/7/8-9

8 Sample889b② (B. breviesepala), 875a③ (B. aristatoserrulata), CP93 (B. ravenii), CP99 (B. hayatana), CP104 (B. breviesepala), CP113 (B. schaaliae), CP153 (B. mingetsensis), CP149 (B. mingetsensis) [CP149太髒,最後回溶再過colum時,溶不掉,又有步驟上的錯誤。]

DNA extraction:CTab (未秤重,目視約兩片正常大小葉子,揉碎直接用鋼珠打碎。) 在65度時放入搖晃機中,約莫一小時,此次只用震盪機 (50 1/s*3 mins)*3 (一顆大、兩顆中鋼珠),開始前要將葉子先撥碎。總體積用50 (-1) ug去溶。皆有過Colum。

NanoDrop (曲老師實驗室)889b② (B. breviesepala):308.8 ug/mL, 1.89 260/280, 2.28 260/230, 875a③ (B. aristatoserrulata):169.6 ug/mL, 1.95 260/280, 2.7 260/230, CP93 (B. ravenii):84.6 ug/mL, 1.85 260/280, 2.02 260/230, CP99 (B. hayatana):81.8 ug/mL, 1.85 260/280, 2.54 260/230, CP104 (B. breviesepala):69.5 ug/mL, 1.82 260/280, 2.09 260/230, CP113 (B. schaaliae):- ug/mL, – 260/280, – 260/230, CP153 (B. mingetsensis):95.4 ug/mL, 1.87 260/280, 2.4 260/230, CP149 (B. mingetsensis):- ug/mL, – 260/280, – 260/230。[波後都有點小崎嶇。]

Note:C用完了,要配藥了。我的心頭好花蓮小檗,這次怎麼沒有出來呢?有夠想你欸!Brenden學長倫太文山那批,不知為何保存的很好 (已詢問保存方法和狀態)。繼續加油。暑假預計8/17-8/25去日本九州,也找學長爬山。

【Experimental Log】2015/7/6-7

8 Sample:b④ (B. breviesepala), 890 (B. breviesepala), 887 (B. kawakamii), CP102 (B. kawakamii), CP144 (B. chingshuiensis), CP145 (B. chingshuiensis), CP148 (B. mingetsensis), CP150 (B. mingetsensis) [CP150有不小心打翻…]

DNA extraction:CTab (未秤重,目視約兩片正常大小葉子,揉碎直接用鋼珠打碎。) 在65度時放入搖晃機中,約莫一小時,此次只用震盪機 (50 1/s*3 mins)*3 (一顆大、兩顆中、兩顆小鋼珠),開始前要將葉子先撥碎。總體積用50 (-1) ug去溶 (887, 890 -3)。皆有過Colum。

NanoDrop (曲老師實驗室)b④ (B. breviesepala)292.4 ug/mL, 2.16 260/280, 2.31 260/230, 890 (B. breviesepala)882.8 (1297.5)(1303.9) ug/mL, 2.12 (2.13)(2.13) 260/280, 2.43 (2.31)(2.3) 260/230, 887 (B. kawakamii)1288.9 (904.2)(885.9) ug/mL, 2.14 (2.14)(2.04) 260/280, 2.35 (2.31)(2.13) 260/230, CP102 (B. kawakamii)730.5 ug/mL, 2.11 260/280, 2.33 260/230, CP144 (B. chingshuiensis)116.4 ug/mL, 2.09 260/280, 1.88 260/230, CP145 (B. chingshuiensis):- ug/mL, – 260/280, – 260/230, CP148 (B. mingetsensis)51.6 ug/mL, 1.83 260/280, 2.16 260/230, CP150 (B. mingetsensis)48.9 ug/mL, 1.76 260/280, 1.57 260/230。[CP148, CP150 大波後有小波。]

Note:最近就專攻高山小檗,有越來越好的趨勢,887和890濃度好高,我濃度測了三次才確定。一直沒有太注意其他的,才發現眠月小檗很壞欸,趁我忙別人偷陰我,葉薄薄,保存看起來都微黑微黑,濃度也不那麼高,可能要快點抽剩下的sample了。台灣小檗依然深得我心 <3。這次抽的CP145是冰冰箱的,從之前到現在,發現加矽膠放冰箱的sample,雖然看起來超綠 (感覺很優),但根本沒用,裏頭幾乎沒有DNA了 (學長說可能因為裏頭水分根本沒乾,DNA就被吃完了),所以冰sample包存法,宣告失敗。

7/5 學長帶回來四棵疑似新種的小檗,希望能好好活著。

7/7 學長出發日本了,祝福他一切順利 !!!

【Experimental Log】2015/6/29-30

8 Sample:895 (B. breviesepala), 896 (B. breviesepala), 897 (B. breviesepala), CP122 (B. nantoensis), CP138 (B. chingshuiensis), CP139 (B. chingshuiensis), CP151 (B. mingetsensis), CP152 (B. mingetsensis)

DNA extraction:CTab (897, CP138量0.2 g, 其餘約在0.1 g左右。) 在65度時放入搖晃機中,約莫一小時,此次只用震盪機 (50 1/s*3 mins)*3 (一顆大、兩顆小鋼珠),開始前要將葉子先撥碎。總體積用50 (-1) ug去溶。皆有過Colum。

NanoDrop (曲老師實驗室)895 (B. breviesepala)149.3 ug/mL,1.84 260/280, 2.11 260/230, 896 (B. breviesepala)23.8 ug/mL, 1.94 260/280, 2.99 260/230, 897 (B. breviesepala)38.8 ug/mL, 1.93 260/280, 2.20 260/230, CP122 (B. nantoensis)18 ug/mL, 1.99 260/280, 5.62 260/230, CP138 (B. chingshuiensis):- ug/mL, – 260/280, – 260/230, CP139 (B. chingshuiensis)50 ug/mL, 1.19 260/280, 0.64 260/230, CP151 (B. mingetsensis)25.8 ug/mL, 1.43 260/280, 1.09 260/230, CP152 (B. mingetsensis)20.1 ug/mL, 2.03 260/280, 2.27 260/230。[895 OK896, 897 大波後面有小波;其餘波醜;CP139不行。]

Note:有點糟。把學長那批高山小檗抽一抽,希望高山小檗總量夠。