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急问：how to dissolve RNA pellets
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急问：how to dissolve RNA pellets# Biology - 生物学
s*t
1 楼
◎译　　名　绿灯侠：绿色骑士/绿灯侠：翡翠骑士
◎片　　名　Green Lantern Emerald Knights
◎年　　代　2011
◎国　　家　美国
◎类　　别　动画/动作/科幻/冒险/幻想
◎语　　言　英语
◎字　　幕　中英双字
◎IMDB评分 7.2/10 (44 votes)
◎IMDB链接 http://www.imdb.com/title/tt1683043
◎文件格式　DVD-RMVB
◎视频尺寸　640 x 352
◎文件大小　1CD 434MB
◎片　　长　84 Mins
◎导　　演　Chris Berkeley
Lauren Montgomery
◎主　　演　Nathan Fillion ... Hal Jordan (voice)
Jason Isaacs ... Sinestro (voice)
Elisabeth Moss ... Arisia (voice)
Kelly Hu ... Laira (voice)
Arnold Vosloo ... Abin Sur (voice)
Steve Blum ... Guardian of OA (voice)
Henry Rollins ... Kilowog (voice)
Roddy Piper ... Bolphunga (voice)
Bruce Thomas ... Atrocitus (voice)
Wade Williams ... Deegan (voice)
David Kaufman ... Ruben (voice)
Sunil Malhotra ... Ship (voice)
◎简　　介
电影讲述绿灯军团面临一次严峻的斗争。　As the home planet of the Green
Lantern Corps faces a battle with an ancient enemy, Hal Jordan prepares new
recruit Arisia for the coming conflict by relating stories of the first
Green Lantern and several of Hal's comrades
下载链接：http://www.filesonic.com/file/1228759884/绿灯侠_绿色骑士.rmvb
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c*e
2 楼
【 以下文字转载自 Joke 讨论区 】
发信人: platinumegg (蛋,纯铂金的), 信区: Joke
标 题: 本周最佳对联
发信站: BBS 未名空间站 (Thu Mar 7 18:21:33 2013, 美东)
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T*d
3 楼
俺装的是双系统，现在系统崩溃了，需要重新装，请教一下如何格式化后重新装。
多谢！
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O*e
4 楼
Those are total RNA isolated from macrophages. I used TE buffer to try to
dissolve the RNA pellets. But the pellets don't go to solution even after I
tried to heat them up to 56 degree. Does anybody have a better idea? Thanks
a lot!
avatar
y*l
5 楼
还敢下？
前段时间不是有两万多个IP被告了
要下得去star bucks或者麦当劳

【在 s*****t 的大作中提到】
: ◎译　　名　绿灯侠：绿色骑士/绿灯侠：翡翠骑士
: ◎片　　名　Green Lantern Emerald Knights
: ◎年　　代　2011
: ◎国　　家　美国
: ◎类　　别　动画/动作/科幻/冒险/幻想
: ◎语　　言　英语
: ◎字　　幕　中英双字
: ◎IMDB评分 7.2/10 (44 votes)
: ◎IMDB链接 http://www.imdb.com/title/tt1683043
: ◎文件格式　DVD-RMVB
avatar
m*o
6 楼
LOL

【在 c********e 的大作中提到】
: 【 以下文字转载自 Joke 讨论区 】
: 发信人: platinumegg (蛋,纯铂金的), 信区: Joke
: 标 题: 本周最佳对联
: 发信站: BBS 未名空间站 (Thu Mar 7 18:21:33 2013, 美东)
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P*a
7 楼
插入随机盘启动，一步一步照屏幕提示做就好了

【在 T*********d 的大作中提到】
: 俺装的是双系统，现在系统崩溃了，需要重新装，请教一下如何格式化后重新装。
: 多谢！
avatar
S*s
8 楼
How did you precipitate RNA? If you use ethanol wash, did you get rid of
ethanol completely? I have tried both LiCl and Ethanol precipitation, then
resolve in either pure water or TE, no problem at all.

I
Thanks

【在 O*********e 的大作中提到】
: Those are total RNA isolated from macrophages. I used TE buffer to try to
: dissolve the RNA pellets. But the pellets don't go to solution even after I
: tried to heat them up to 56 degree. Does anybody have a better idea? Thanks
: a lot!
avatar
c*e
9 楼
pc很识货。

【在 m*******o 的大作中提到】
: LOL
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T*d
10 楼
多谢答复，现在问题是插入随机盘后，提示要装在那个区域，而又没有显示出来那个区
域，
所以第一步要格式化，俺不知道如何格式化。

【在 P***a 的大作中提到】
: 插入随机盘启动，一步一步照屏幕提示做就好了
avatar
O*e
11 楼
It was ethanol precipitation, 100% first and then wash with 70%
I've been dealing with RNA for quite some time, never had this problem
before.
The RNA was from peritoneal macrophages. I was warned before about the tough
RNA pellets, and just realized it was such a pain ...

【在 S*****s 的大作中提到】
: How did you precipitate RNA? If you use ethanol wash, did you get rid of
: ethanol completely? I have tried both LiCl and Ethanol precipitation, then
: resolve in either pure water or TE, no problem at all.
:
: I
: Thanks
avatar
w*e
12 楼
how did you isolate the RNA? trizol? did you use enough? is the pellet
oily?
I had similar problem before.

I
Thanks

【在 O*********e 的大作中提到】
: Those are total RNA isolated from macrophages. I used TE buffer to try to
: dissolve the RNA pellets. But the pellets don't go to solution even after I
: tried to heat them up to 56 degree. Does anybody have a better idea? Thanks
: a lot!
avatar
a*k
13 楼
don't let the RNA pellet dry completely, it'll greatly decrease its
solubility

I
Thanks

【在 O*********e 的大作中提到】
: Those are total RNA isolated from macrophages. I used TE buffer to try to
: dissolve the RNA pellets. But the pellets don't go to solution even after I
: tried to heat them up to 56 degree. Does anybody have a better idea? Thanks
: a lot!
avatar
O*e
14 楼
trizol
i think I used plenty
I can see the pellets floating in the TE, not oily though
I did the isolation with trizol samples from other cells, they were just
fine (nice pellets, dissolved easily in TE), so I'm thinking it's the
macrophage RNA that gives me trouble.
anybody on this board working with macrophages?

【在 w******e 的大作中提到】
: how did you isolate the RNA? trizol? did you use enough? is the pellet
: oily?
: I had similar problem before.
:
: I
: Thanks
avatar
w*a
15 楼
Macrophages seem to be different from other cell types. I have exactly
the same experience before. Eventully, I switched to Qiagen RNeasy Kit
for RNA extraction. The yield is lower, while the RNA quality is good enough.

【在 O*********e 的大作中提到】
: trizol
: i think I used plenty
: I can see the pellets floating in the TE, not oily though
: I did the isolation with trizol samples from other cells, they were just
: fine (nice pellets, dissolved easily in TE), so I'm thinking it's the
: macrophage RNA that gives me trouble.
: anybody on this board working with macrophages?
avatar
n*w
16 楼
干得太透了吧。。。
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a*i
17 楼
I only use water to dissolve.....then it disappears
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O*e
18 楼
天，这个都会上首页。。
多谢wdgca的回复，我也觉得是macrophage的问题。下次用那个kit试试看
不是干透的问题，我提RNA已经有4年了，从来不用speed vacuum，都是风干的，pellet
一变透明就加TE。TE和water我都用过，做microarray的样品用nuclease-free water，
其他没那么高质量要求的时候就用TE，没发现什么区别。还有我前面说过，我同时提的
macrophage和splenocyte的RNA，后者一点问题都没有，macrophage的RNA pellet其实
还小些，但就是不溶。昨天折腾了好久，还是能看到薄薄的一片pellet飘在管子里，但
测 OD到了100ng/ul，足够一个反转录PCR了,就将就着用了。下次一定试试那个Qiagen
RNeasy Kit。也给以后要提macrophage RNA的同学提个醒，really tough pellet，要
是样品宝贵的话不要自己提，直接上kit。
avatar
Y*I
19 楼
vortex到手酸。
俺曾经vortex RNA pellet40分钟。。。最终被击破。

pellet
Qiagen

【在 O*********e 的大作中提到】
: 天，这个都会上首页。。
: 多谢wdgca的回复，我也觉得是macrophage的问题。下次用那个kit试试看
: 不是干透的问题，我提RNA已经有4年了，从来不用speed vacuum，都是风干的，pellet
: 一变透明就加TE。TE和water我都用过，做microarray的样品用nuclease-free water，
: 其他没那么高质量要求的时候就用TE，没发现什么区别。还有我前面说过，我同时提的
: macrophage和splenocyte的RNA，后者一点问题都没有，macrophage的RNA pellet其实
: 还小些，但就是不溶。昨天折腾了好久，还是能看到薄薄的一片pellet飘在管子里，但
: 测 OD到了100ng/ul，足够一个反转录PCR了,就将就着用了。下次一定试试那个Qiagen
: RNeasy Kit。也给以后要提macrophage RNA的同学提个醒，really tough pellet，要
: 是样品宝贵的话不要自己提，直接上kit。
avatar
O*e
20 楼
pfpf~ 哈哈

【在 Y**I 的大作中提到】
: vortex到手酸。
: 俺曾经vortex RNA pellet40分钟。。。最终被击破。
:
: pellet
: Qiagen
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