H*****e
发帖数: 120 | 1 Hi experts,
I need to sequence PI3K and PTEN in tumors for determine whether there is
mutation. But I do not know much about sequencing mutations since I only
work kinase for years. So I have a few basic questions, wish to get answers
here:
1.RNA or DNA, The tumor sample will be provided by a colleague in Hospital.
I can get both DNA and RNA purified. Should I sequence DNA or RNA after I
PCR them out?
2.If only 1 copy has mutation and the other copy is wt, can routine sequence
in core lab detect it? Or I have to use the so-called pyrosequence?
Thanks! | k*******s
发帖数: 214 | 2 The fast way is to PCR the target region and sequence. If only one mutation
, there will be two peaks at the specific sit and it can seen from the
sequencing chromatograph.
answers
.
I
sequence
【在 H*****e 的大作中提到】
: Hi experts,
: I need to sequence PI3K and PTEN in tumors for determine whether there is
: mutation. But I do not know much about sequencing mutations since I only
: work kinase for years. So I have a few basic questions, wish to get answers
: here:
: 1.RNA or DNA, The tumor sample will be provided by a colleague in Hospital.
: I can get both DNA and RNA purified. Should I sequence DNA or RNA after I
: PCR them out?
: 2.If only 1 copy has mutation and the other copy is wt, can routine sequence
: in core lab detect it? Or I have to use the so-called pyrosequence?
| O******e
发帖数: 4845 | 3 1. I would get both DNA and RNA, just in case RNA-Seq becomes an option in
the future.
2. Yes.
answers
.
I
sequence
【在 H*****e 的大作中提到】
: Hi experts,
: I need to sequence PI3K and PTEN in tumors for determine whether there is
: mutation. But I do not know much about sequencing mutations since I only
: work kinase for years. So I have a few basic questions, wish to get answers
: here:
: 1.RNA or DNA, The tumor sample will be provided by a colleague in Hospital.
: I can get both DNA and RNA purified. Should I sequence DNA or RNA after I
: PCR them out?
: 2.If only 1 copy has mutation and the other copy is wt, can routine sequence
: in core lab detect it? Or I have to use the so-called pyrosequence?
| l**********1
发帖数: 5204 | 4 LZ you can try
NGS 基因测序仪IonProton
References:
a)
“个人基因组测序”时代来临 只需1000美元
2012年04月01日
来源：新华网
总部位于美国加州的生命科学厂商LifeTechnologies公司，29日宣布在中国推出新的台式基因测序仪IonProton。借助
该技术产品，只需1000美元即可在一天时间内完成个人全基因组测序。这意味着基因测序技术有望走进临床实践，“飞
入寻常百姓家
//tech.ifeng.com/discovery/detail_2012_04/01/13603299_0.shtml
b)
NGS SNP analysis Software:
for example:
V-Phaser & V-Profiler: variant calling a visualization tools to identify
biological mutations in diverse populations
//www.broadinstitute.org/scientific-community/science/projects/viral-
genomics/v-phaser
//www.broadinstitute.org/scientific-community/software?criteria=Next%
20generation%20tools
//www.broadinstitute.org/scientific-community/software?criteria=RNA-seq
more try
//www.broadinstitute.org/scientific-community/software
and
//www.broadinstitute.org/scientific-community/data
//www.broadinstitute.org/
all above links are from:
//www.shigen.nig.ac.jp/rice/oryzabase/tool/tool.jsp;jsessionid=
0D7767EEC6B68517E0FDE8DDAA20F6A9.4_6
【在 O******e 的大作中提到】
: 1. I would get both DNA and RNA, just in case RNA-Seq becomes an option in
: the future.
: 2. Yes.
:
: answers
: .
: I
: sequence
|
|
