chip实验求教：大家用0.5%的SDS还是用1%的SDS - 未名空间MITBBS历史存档

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chip实验求教：大家用0.5%的SDS还是用1%的SDS

chip实验求教：大家用0.5%的SDS还是用1%的SDS# Biology - 生物学

p*w2012-12-10 08:12

1 楼

漂亮小MM泪眼汪汪求二三流艺术家的瓜子（图）zz

t*r2012-12-10 08:12

2 楼

可是只有我的case还是查不到。
我是3月初递的，有类似情况吗？
上星期把指纹打过了。
★ Sent from iPhone App: iReader Mitbbs Lite 7.28

t*82012-12-10 08:12

3 楼

我现在用的protocol，是crosslinking后用0.5%的SDS lysis cells。我见网上很多
protocol都是用1%的SDS，因为我的蛋白可能比较难抽，我就改用1%的SDS做，结果比0.
5%的SDS回收率还低。大家一般是用哪个浓度啊？谢谢！

c*n2012-12-10 08:12

4 楼

俺也是3月初的，除了FP还啥都没有呢。。。。。。

【在 t*****r 的大作中提到】

: 可是只有我的case还是查不到。
: 我是3月初递的，有类似情况吗？
: 上星期把指纹打过了。
: ★ Sent from iPhone App: iReader Mitbbs Lite 7.28

k*n2012-12-10 08:12

5 楼

我们用的是rick young 的protocol。
Lysis Buffer 1 Add protease inhibitors just before use, filter and keep cold
. Consists of 50 mM HEPES-KOH, pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol,
0.5% NP-40, 0.25% Triton X-100, 1× protease inhibitors Lysis Buffer 2 Add
protease inhibitors just before use, filter and keep cold. Consists of 10 mM
Tris-HCl, pH 8.0, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 1× protease
inhibitors
Lysis Buffer 3 Add protease inhibitors just before use, filter and keep cold
. Consists of 10 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0
.1% Na-Deoxycholate, 0.5% N-lauroylsarcosine, 1× protease inhibitors

z*o2012-12-10 08:12

6 楼

俺也是，
看到TSC3月初交的，大部分都批了
bless us
有消息了通知一声

t*82012-12-10 08:12

7 楼

谢谢你的热心回复。你冒似用的是两步裂解。你最后用的是RIPA buffer,不是SDS。我
的蛋白不用SDS可能抽不出来，我决定还是用1％SDS吧

cold
glycerol,
Add
mM
cold
0

【在 k*****n 的大作中提到】

: 我们用的是rick young 的protocol。
: Lysis Buffer 1 Add protease inhibitors just before use, filter and keep cold
: . Consists of 50 mM HEPES-KOH, pH 7.5, 140 mM NaCl, 1 mM EDTA, 10% glycerol,
: 0.5% NP-40, 0.25% Triton X-100, 1× protease inhibitors Lysis Buffer 2 Add
: protease inhibitors just before use, filter and keep cold. Consists of 10 mM
: Tris-HCl, pH 8.0, 200 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 1× protease
: inhibitors
: Lysis Buffer 3 Add protease inhibitors just before use, filter and keep cold
: . Consists of 10 mM Tris-HCl, pH 8.0, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0
: .1% Na-Deoxycholate, 0.5% N-lauroylsarcosine, 1× protease inhibitors

x*g2012-12-10 08:12

8 楼

You can actually call to ask if there is any issue with your case. I called
twice and was told my case is fine before I finally got my approval notice,
which is much slower than number around me. They told me that my security
check takes more time.

m*v2012-12-10 08:12

9 楼

Would you please tell us the phone # and options you selected, thanks.

called
,

【在 x**********g 的大作中提到】

: You can actually call to ask if there is any issue with your case. I called
: twice and was told my case is fine before I finally got my approval notice,
: which is much slower than number around me. They told me that my security
: check takes more time.

x*g2012-12-10 08:12

10 楼

there is a 800 number on the uscis check status page.
I don't remember the exact options I selected but there is one step which
will ask if you have a case number (or tracking number). Choose no at that
step and you will hear some instruction regarding how to find your case
number and continue choose no and you will be directed to a representative
who won't know anything. Just kindly tell him/her your case(e.g. I told them
that I am the dependent and my wife's case has been approved for the second
call), they will direct you to someone who can actually access your case,
have your A# ready and you can ask whatever you want, but be polite.
PM me if you have any questions.

【在 m******v 的大作中提到】

: Would you please tell us the phone # and options you selected, thanks.
:
: called
: ,