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2005年12月02日

minasan konbanwa.

its almost 3:30am in ca...and im still working on my baby clones. couple days ago, i mini-preped the bac, yet it turned out to show almost nothing on a260. i thought i somehow got messed with prepping, then went for another prep. however, however, however, i got the same stupid a260 again. it shouldnt be possible, so now im running agarose just to check plasmid IS in the preped samples. if the spec is found to have a problem, which is probably what it is, that means i wasted last few weeks since ive been having a hard time to get enough plasmid to finishing up virus construction.
first time i tried to clone them, i just picked dh5-alpha with an easy thought it would do good enough. actually, agarose showed a strong band for the digested samples while a260 ended up with pretty low conc of double strand dna. that was the time i explored and found that dh5-alpha is not suitable for lenti vector. then i walked around other labs and got other two strains of stbl4 and exlt blue. they grew up well on amp, meaning they should have gotten the plasmid. but, a260 is saying there is no dna (or even any possible contamination). its so wierd...i have no idea what to do if there is not any problem with the spec machine. hmmm...well...it will be envailed in at least a half hour anyway.
i guess everyone has been busy these days but hope everything will go great with all of you guys. those in the states, have a sweet dream. those back in japan, have a good rest. poke me if you are bored.

good nite.

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最終更新日 2005年12月02日 22時37分18秒
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Jaguar XJ6 さん

やっぱりspecのせいでした。

1回目のmini-prepのときも真夜中で半分寝ぼけながらやっとったけん、何か「途中でeluteしたんかなー」とか思っとったとけど、んなことはなかったです。
しかも、1回目のは最後もelution bufferでやらずに水でeluteしたけん「まぁ途中で流したとならblankとまったく同じabsorbanceでも当たり前よね。」とか納得してしまった。
でも眠くて腹も立ちません。
結局今日も朝の6時。朝のクラスまで少し寝まーす。
おやすみなさい。 (2005年12月03日 19時13分27秒)

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