Salmon Sperm DNA/Protein A Agarose, Catalog #16-157C, Lot # 0608038187. One vial containing 600μg sonicated Salmon Sperm DNA, 1.5mg BSA and approximately 4.5 mg recombinant Protein A. Provided as a 50% gel slurry for a final volume of 3mL. Suspended in TE Buffer containing 0.05% sodium azide.
SDS Lysis Buffer, Catalog # 20-163. One vial containing 10 ml of 1% SDS, 10 mM EDTA, 50 mM Tris-HCl, pH 8.1.
ChIP Dilution Buffer, Catalog # 20-153. Two vials, each containing 24 mL of 0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.1, 167 mM NaCl.
Low Salt Immune Complex Wash Buffer, Catalog #20-154. One vial containing 24 mL of 0.1% SDS, 1%Triton X-100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 150mM NaCl.
High Salt Immune Complex Wash Buffer,Catalog # 20-155. One vial containing 24 mL of 0.1% SDS, 1% Triton X-100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 500 mM NaCl.
LiCl Immune Complex Wash Buffer, Catalog #20-156. One vial containing 24 mL of 0.25 M LiCl, 1% NP40, 1% deoxycholate, 1 mM EDTA, 10 mM Tris-HCl, pH 8.1.
TE Buffer, Catalog # 20-157. Two vials, each containing 24 mL of 10 mM Tris-HCl, 1 mM EDTA, pH 8.0.
5 M NaCl, Catalog # 20-159. One vial containing 500 μL of 5 M NaCl.
0.5 M EDTA, Catalog # 20-158. One vial containing 250 μL of 0.5M EDTA, pH 8.0.
1 M Tris-HCl, pH 6.5, Catalog # 20-160. One vial containing 500 μL of 1 M Tris-HCl, pH 6.5.
超声断裂DNA是吗？不知道你用的什么型号的超声仪，我用的是Branson 450，我一般设置是这样：
10%的amplitude，
turn sonifier on 0.5 second then off 0.5 second--20 cycles,
then put the sample on ice for a while, repeat for another 3-4 times ( another 3-4 times of the 20 cycles).
烈完后，短时离心，取上清3微升电泳一下看看
gogo (2013-6-27 15:21:02)
QUOTE:
原帖由 fei1226com 于 2013-6-27 15:20 发表
超声断裂DNA是吗？不知道你用的什么型号的超声仪，我用的是Branson 450，我一般设置是这样：
10%的amplitude，
turn sonifier on 0.5 second then off 0.5 second--20 cycles,
then put the sample on ice for a while, re ...
最新回复
fei1226com (2013-6-27 11:27:41)
附上我的CHIP （vendor: upstate）所用试剂盒提供的说明，大多溶液都可以自己配，钱不多的话可以考虑
Salmon Sperm DNA/Protein A Agarose, Catalog #16-157C, Lot # 0608038187. One vial containing 600μg sonicated Salmon Sperm DNA, 1.5mg BSA and approximately 4.5 mg recombinant Protein A. Provided as a 50% gel slurry for a final volume of 3mL. Suspended in TE Buffer containing 0.05% sodium azide.
SDS Lysis Buffer, Catalog # 20-163. One vial containing 10 ml of 1% SDS, 10 mM EDTA, 50 mM Tris-HCl, pH 8.1.
ChIP Dilution Buffer, Catalog # 20-153. Two vials, each containing 24 mL of 0.01% SDS, 1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl, pH 8.1, 167 mM NaCl.
Low Salt Immune Complex Wash Buffer, Catalog #20-154. One vial containing 24 mL of 0.1% SDS, 1%Triton X-100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 150mM NaCl.
High Salt Immune Complex Wash Buffer,Catalog # 20-155. One vial containing 24 mL of 0.1% SDS, 1% Triton X-100, 2 mM EDTA, 20 mM Tris-HCl, pH 8.1, 500 mM NaCl.
LiCl Immune Complex Wash Buffer, Catalog #20-156. One vial containing 24 mL of 0.25 M LiCl, 1% NP40, 1% deoxycholate, 1 mM EDTA, 10 mM Tris-HCl, pH 8.1.
TE Buffer, Catalog # 20-157. Two vials, each containing 24 mL of 10 mM Tris-HCl, 1 mM EDTA, pH 8.0.
5 M NaCl, Catalog # 20-159. One vial containing 500 μL of 5 M NaCl.
0.5 M EDTA, Catalog # 20-158. One vial containing 250 μL of 0.5M EDTA, pH 8.0.
1 M Tris-HCl, pH 6.5, Catalog # 20-160. One vial containing 500 μL of 1 M Tris-HCl, pH 6.5.
sr9971 (2013-6-27 11:28:07)
谢谢。看来试剂还挺多的，估计不是这么容易凑齐的。你买这个试剂盒花了多少银子啊。
fei1226com (2013-6-27 11:28:36)
QUOTE:
不客气，花了464美刀。可以做22次。其实上面好几个溶液都是比较常用的东西，我们实验室的人做CHIP就是自己配，因为我是接下来要做CHIP-on-chip，害怕浪费片子，保险起见，所以就用了现成的试剂盒。
如果自己配溶液，注意最后过滤一下，昨天忘了说了
flower-201 (2013-6-27 15:18:07)
yonger (2013-6-27 15:18:31)
相关疾病：
肿瘤
你在那里哪?我是河北的博士,想做肿瘤的Chip,但我们这里的实验室不能做.不知哪里的实验室可做此实验,我可出费用.我的手机:13230161095
fei1226com (2013-6-27 15:19:03)
QUOTE:
不好意思我现在国外。你们实验室能做pcr或者wb吗？如果能做的话，chip应该是没有问题的，就是试剂的问题，我可以把我的protocol发给你。如果你要做ChIP-on-chip，估计要联系公司了wmp1234 (2013-6-27 15:19:25)
fei1226com (2013-6-27 15:19:51)
QUOTE:
你好，我用的是agilent的244k， human CpG island microarrayzzzz (2013-6-27 15:20:12)
你最好自己买试剂盒，配试剂很累的，而且一个pH就可能影响你的结果，我就超声都一个多月了，还没有一个理想结果。超声太压抑了，费力不讨好！
fei1226com (2013-6-27 15:20:36)
QUOTE:
超声断裂DNA是吗？不知道你用的什么型号的超声仪，我用的是Branson 450，我一般设置是这样：10%的amplitude，
turn sonifier on 0.5 second then off 0.5 second--20 cycles,
then put the sample on ice for a while, repeat for another 3-4 times ( another 3-4 times of the 20 cycles).
烈完后，短时离心，取上清3微升电泳一下看看
gogo (2013-6-27 15:21:02)
QUOTE:
直接电泳检测超声效果可以吗？我看很多prptocol上都要解交联，抽提后再电泳分析。fei1226com (2013-6-27 15:21:26)
QUOTE:
可以我就是这么做的，电泳前全速离一下心，去上清跑电泳就行，我开始也是因为懒得reverse crosslink，因为一做就要4个小时，耽误不起啊，万一裂解不够，还得重来过，太浪费时间。我这么做了好几次了，没有问题TAT (2013-6-27 15:21:45)
【求助】请教关于CHIP的问题