【Experimental Log】2016/03/28

17 Samples:CP227 (B. alpicola), CPL15 (B. aristatoserrulata), CPL16 (B. aristatoserrulata), CP191 (B. breviesepala), CPL18 (B. breviesepala), CP221 (B. kawakamii), CP214 (B. kawakamii), CPL10 (B. ravenii), CPL11 (B. ravenii), CPL12 (B. ravenii), CPL3 (B. nantoensis), CP231-1 (B. pengii), CP232-1 (B. pengii), CPL13 (B. pengii), CPL8 (B. tarokoensis), CPL9 (B. tarokoensis), CPL4 (B. mingetsensis)

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。[CPL10 (B. ravenii), CPL11 (B. ravenii), CPL12 (B. ravenii)]

DNA定量:Qbit [CPL10 (B. ravenii), CPL11 (B. ravenii), CPL12 (B. ravenii), CPL15 (B. aristatoserrulata),由於在20160223抽的那支,濃度too high,所以這次稀釋一半後再測濃度。]

Qbit結果:CPL15 (B. aristatoserrulata):69.4 ng/uL, CPL10 (B. ravenii):50.2 ng/uL, CPL11 (B. ravenii):59.2 ng/uL, CPL12 (B. ravenii):too high

跑膠:CP227 (B. alpicola), CPL15 (B. aristatoserrulata), CPL16 (B. aristatoserrulata), CP191 (B. breviesepala), CPL18 (B. breviesepala), CP221 (B. kawakamii), CP214 (B. kawakamii), CPL10 (B. ravenii), CPL11 (B. ravenii), CPL12 (B. ravenii), CPL3 (B. nantoensis), CP231-1 (B. pengii), CP232-1 (B. pengii), CPL13 (B. pengii), CPL8 (B. tarokoensis), CPL9 (B. tarokoensis), CPL4 (B. mingetsensis)

20160328跑膠

結果:跑膠結果皆OK!

廣告

【Experimental Log】2016/03/19

10 Samples:CPL6 (B. breviesepala), CP221 (B. hayatana), CP237-1 (B. kawakamii), CP117 (B. ravenii), CP181 (B. ravenii), CP132 (B. schaaliae), B318a (B. baradana), 武1 (B. wuyiensis), 武2 (B. wuyiensis), 武3 (B. wuyiensis)

純化kit:Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB) [CP117 (B. ravenii), CP181 (B. ravenii), 武1 (B. wuyiensis), 武2 (B. wuyiensis), 武3 (B. wuyiensis),原來CP117和CP181只是要跑膠,赫然發現這兩支沒有過純化,原液顏色汙濁,故立馬過clean up,在測Qbit。]

DNA定量:Qbit [CPL6 (B. breviesepala), CP237-1 (B. kawakamii), B318a (B. baradana), 武1 (B. wuyiensis), 武2 (B. wuyiensis), 武3 (B. wuyiensis), CP117 (B. ravenii), CP181 (B. ravenii)]

Qbit結果:CPL6 (B. breviesepala):57.2 ng/uL, CP237-1 (B. kawakamii):68.8(69.4) ng/uL, B318a (B. baradana):56.8 ng/uL, 武1 (B. wuyiensis):38.6 ng/uL, 武2 (B. wuyiensis):34.4 ng/uL, 武3 (B. wuyiensis):55 ng/uL, CP117 (B. ravenii):21.6 ng/uL, CP181 (B. ravenii):12.4 ng/uL

跑膠結果:[CPL6 (B. breviesepala), CP221 (B. hayatana), CP132 (B. schaaliae), 武1 (B. wuyiensis), 武2 (B. wuyiensis), 武3 (B. wuyiensis)]

20160319跑膠

結論:跑膠OK!但是但是但是但是,非常可怕的漏網之魚→CP117 (B. ravenii):21.6 ng/uL, CP181 (B. ravenii):12.4 ng/uL,原來在20151022測得77.2和66.7!濃度都是髒東西,兩支皆不能使用,過完Cleanup才發現,濃度不夠!緊急重抽!(幸好還有活體!)

【Experimental Log】2016/03/24

6 Samples:M1 (B. mori), CP238-1 (B. pengii), CP233-1 (B. pengii), CP224 (B. kawakamii), CP123 (B. nantoensis), CP141 (B. chingshuiensis)

稀釋 CP238-1 (B. pengii), CP233-1 (B. pengii), CP224 (B. kawakamii從原液中取25uL,再加25uL稀釋。因為原本為測得濃度為too high。

DNA定量:Qbit

Qbit結果:CP238-1 (B. pengii):ˊ66(66.8) ng/uL, CP233-1 (B. pengii):58 ng/uL, CP224 (B. kawakamii) :60 ng/uL。

跑膠:M1 (B. mori), CP233-1 (B. pengii), CP224 (B. kawakamii), CP123 (B. nantoensis), CP141 (B. chingshuiensis)

結論:跑膠OK。發現之前測試為too high的sample,稀釋為兩倍後,濃度都大約再60 ng/uL左右。

已整理出要上機的sample了。

【Experimental Log】2016/03/18-19

6 Samples:B318a (B. baradana), B318b (B. baradana), CP102 (B. kawakamii), CP124 (B. kawakamii), CP142 (B. chingshuiensis), CP184 (B. nantoensis)

DNA extraction:CTab (為乾燥材料,均質機打碎。半片葉子一管,一片葉子就兩管 (如B318a-1和B318a-2),最後將兩管濃縮成一管。) 總體積用50 ug去溶。[B318a & B318b,為英國爺爺寄給我的菲律賓小檗,20160318抵達。]

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

DNA定量:Qbit

Qbit結果:B318a (B. baradana):too high ng/uL, B318b (B. baradana):102 ng/uL。

跑膠結果[B318a (B. baradana), B318b (B. baradana), CP102 (B. kawakamii), CP124 (B. kawakamii), CP142 (B. chingshuiensis), CP184 (B. nantoensis)]

20160319跑膠

結論:發現新收到的B318a & B318b B. baradana ,能用!其他的(CP系列)片段大小都還算大。最後的菲律賓小檗也 (終於) 收齊了。

【Experimental Log】2016/03/10

8 Samples:D1 (B. baradana), D5 (B. baradana), D6 (B. baradana), D7 (B. baradana), CP104 (B. breviesepala), CP170 (B. breviesepala), CP173 (B. hayatana), 745 (B. aristatoserrulata)

純化kit:Monarch™ PCR & DNA Cleanup Kit (5 μg) (NEB) [過漢堯學長抽的D1, D5, D6, D7,只溶20 μL]

DNA定量:Qbit [漢堯學長抽的D1, D5, D6, D7,最後溶20 μL]

Qbit結果:D1 (B. baradana):73.6 ng/uL, D5 (B. baradana):39.4(39.2) ng/uL, D6 (B. baradana):112 ng/uL, D7 (B. baradana):79.2 ng/uL

跑膠結果

20160310跑膠v1

結論:發現漢堯學長抽的B. baradana D6, D7,似乎還用!(意外的驚喜) 我的(CP系列)片段大小都還算大。學長的sample(745)smear嚴重。

【Experimental Log】2016/03/06

12 Samples:726 (B. aristatoserrulata), CP88 (B. aristatoserrulata), CP139 (B. chingshuiensis), CP99 (B. hayatana), 719 (B. hayatana), CP61 (B. kawakamii), CP172 (B. kawakamii), 709 (B. mingtsuensis), CP115 (B. schaaliae), 887 (B. kawakamii), 765 (B. kawakamii), S11 (B. baradana)

跑膠結果

20160306跑膠v2

結論:和預料中的差不多。(CP系列)片段大小都還算大,學長的Sample還是不行。跑膠沒有想像中可怕啦。

跑膠protocal
大片:25ml, 小片:15ml
膠:1.0%
healthy dye:5ul/100ml
dye=sample*1/6
marker:2~4ul

【Experimental Log】2016/03/02

12 Samples:S5 (B. baradana), S6 (B. baradana), S13 (B. baradana), CP238-1 (B. pengii), 760 (B. schaaliae), 698 (B. breviesepala), CP144 (B. chingshuiensis), CP82 (B. aristatoserrulata), CPL2 (B. nantoensis), 705 (B. mingtsuensis), CP185 (B. tarokoensis), CP92 (B. ravenii)

跑膠結果

20160302跑膠

結論:發現我的(CP系列)片段大小都還算大。學長的sample(760, 698, 705)smear嚴重。必須重新整理,學長之前給我的sample是否必定要使用。因為希望還是希望都像O那樣品質的上機比較好。然後,要開始練習跑膠了。(憂心忡忡)