g******c
发帖数: 18 | 1 Try Ca for DNA experiments.
Try the same experiments shown on S figure 2 by using DNA instead of RNA as
a substrate.
Han Lab's Mg reagent may be contaminated with Ca or other metal cations.
Good luck to all |
d********0
发帖数: 534 | 2 哈哈，真巧，我也这么想过，可能他们的实验用水或其它试剂不合格反而导致做出来了
，可惜自己没条件做，哪位有钱有闲的人员可以自己try一下，保不齐试出来了就快申
请专利 |
l*******1
发帖数: 16217 | 3 韩春雨这个事情发展到现在还没有一个欧美大牛跳出来说话，你看人家都沉得住气 |
l*******1
发帖数: 16217 | 4 欧美在面对这种新生事物诞生时，明显更有经验，更有耐心和宽容，
反观一群小黄人儿是跳梁小丑般的表演 |
w***9
发帖数: 804 | 5 其实我们黄种人做的也很不错，比如我，从一开始就反复说明要尊重科学精神，强调科
学需要宽容。
我是本版唯一没有骂人，没有讲脏话，只是因为支持韩副主席被封的人。但是我从来没
有抱怨任何人。对待反对韩副主席的人，不妨也给他们一点宽容，因为他们没有，最需
要。希望他们拿到宽容以后，以后也能送出去。
【在 l*******1 的大作中提到】
: 欧美在面对这种新生事物诞生时，明显更有经验，更有耐心和宽容，
: 反观一群小黄人儿是跳梁小丑般的表演
|
g******c
发帖数: 18 | 6 PfANgo cuts DNA, by using Mn and Co
NgAgo cuts RNA by using Mn and Mg
Metal Cation size matters.
A few Asp residues are important for the activity of pfAgo, but not for
NgAgo. D residues are normally the binding site of metal cations.
Go figure the native environment of NgAgo and may have clues.
Importantly, NgAgo is active at 37, while other Agos are not. |
m******y
发帖数: 38 | |
N****n
发帖数: 294 | 8 你是证实了二价离子的作用，还是根据模型推理出来的？
【在 g******c 的大作中提到】
: PfANgo cuts DNA, by using Mn and Co
: NgAgo cuts RNA by using Mn and Mg
: Metal Cation size matters.
: A few Asp residues are important for the activity of pfAgo, but not for
: NgAgo. D residues are normally the binding site of metal cations.
: Go figure the native environment of NgAgo and may have clues.
: Importantly, NgAgo is active at 37, while other Agos are not.
|
l*******1
发帖数: 16217 | 9 楼上的生物化学一看就没学好
Cofactors, mostly metal ions or coenzymes, are inorganic and organic
chemicals that assist enzymes during the catalysis of reactions. |
N****n
发帖数: 294 | 10 你说的也对。
不过我关心的是Ca2+一般不在DNA/RNA酶里起作用。Mn2+有可能改变底物专一性
【在 l*******1 的大作中提到】
: 楼上的生物化学一看就没学好
: Cofactors, mostly metal ions or coenzymes, are inorganic and organic
: chemicals that assist enzymes during the catalysis of reactions.
|
g******c
发帖数: 18 | 11 Please consider the inhibition effects caused by high affinity unfunctional
metal cations that compete against the functional metal cations. So it's
important to do extensive dialysis after the NgAgo is purified by affinity
chromatography before one carries out the in vitro nuclease activity assay.
Never know what's going on really .
Han has to figure the reason quickly. If other labs figure it out before
they do, the credit will go to that lab.
: PfANgo cuts DNA, by using Mn and Co
: NgAgo cuts RNA by using Mn and Mg
: Metal Cation size matters.
: A few Asp residues are important for the activity of pfAgo, but not
for
: NgAgo. D residues are normally the binding site of metal cations.
: Go figure the native environment of NgAgo and may have clues.
: Importantly, NgAgo is active at 37, while other Agos are not.
【在 g******c 的大作中提到】
: PfANgo cuts DNA, by using Mn and Co
: NgAgo cuts RNA by using Mn and Mg
: Metal Cation size matters.
: A few Asp residues are important for the activity of pfAgo, but not for
: NgAgo. D residues are normally the binding site of metal cations.
: Go figure the native environment of NgAgo and may have clues.
: Importantly, NgAgo is active at 37, while other Agos are not.
|
m****a
发帖数: 270 | 12 Welcome back. Brother Jianghu. You are the first to suggest EDTA need to be
avoided in Ngago transfection.
You are truly a legend.
unfunctional
【在 g******c 的大作中提到】
: Please consider the inhibition effects caused by high affinity unfunctional
: metal cations that compete against the functional metal cations. So it's
: important to do extensive dialysis after the NgAgo is purified by affinity
: chromatography before one carries out the in vitro nuclease activity assay.
: Never know what's going on really .
: Han has to figure the reason quickly. If other labs figure it out before
: they do, the credit will go to that lab.
:
:
: PfANgo cuts DNA, by using Mn and Co
:
: NgAgo cuts RNA by using Mn and Mg
|
d*******g
发帖数: 8992 | 13 外行的说一句，Ca2+对于肌肉的作用也是二元的，低浓度时激活，高浓度时抑制 |
