FGFR fusion related

2/27(四)合作討論會議: BGJ398 A2204 cholangiocarcinoma study

#1由 sufang 在 三, 04/09/2014 - 12:08 發表。

Norvatis人員的報告摘要

1. K650E in FGFR3 mutation is also sensitive to BGJ398.

2. Dan/YiMi的paper是K652E對PD173074無效。

3. TKI258不知道是啥...有點失憶!

2014-03-04 Journal Club Paper

Dear all:

...20140304 Journal club 輪到我向各位報告。
題目: Hepatocellular Carcinoma-Associated Mesenchymal Stem Cells Promote Hepatocarcinoma Progression: Role of the S100A4-mIR155-SOCS1-MMP9 Axis

Paper 於附檔請查收~~~謝謝大家!!

QPCR validation of gene fusion on clinical samples

由 sufang 在 二, 04/08/2014 - 09:42 發表 Cholangiocarcinoma Dan Robinson MCTP RNA-seq UMich Yi-Mi

For QPCR validations we use real-time PCR with SYBR green on an ABI instrument such as a StepOne, we set the run for 40 cycles, cDNA from 200 ng or so of toral RNA. Using GAPDH control primers we get a Ct of 18 cycles or so.

For a real fusion, the Ct can be anywhere from 24 to 33 but not much above that. We use ABI Fast SYBR Green mastermix kits with UNG for our PCRs. One trouble with the PCR approach is the number of 3’ partners and the number of possible fusion junctions for each 3’ partner. (The FGFR fusion exon is nearly always the same.)

For QPCR validations we use real-time PCR with SYBR green on an ABI instrument such as a StepOne, we set the run for 40 cycles, cDNA from 200 ng or so of toral RNA. Using GAPDH control primers we get a Ct of 18 cycles or so.

For a real fusion, the Ct can be anywhere from 24 to 33 but not much above that. We use ABI Fast SYBR Green mastermix kits with UNG for our PCRs. One trouble with the PCR approach is the number of 3’ partners and the number of possible fusion junctions for each 3’ partner. (The FGFR fusion exon is nearly always the same.)

2014-02-18 Journal Club Paper

Title:Claudin-2 Promotes Breast Cancer Liver Metastasis by Facilitating Tumor Cell Interactions with Hepatocytes by En-Wei
(原文下載)

20140212 Lab meeting - ingrid

LabMeeting-Ingrid

由 EVKVLIN 在 三, 02/12/2014 - 00:00 發表 LIN Lab 2016 Ingrid LabMeeting YCK

LabMeeting-Ingrid 由 EVKVLIN 在 三, 02/12/2014 - 00:00 發表  LIN Lab 2016   Ingrid   LabMeeting   YCK 2008/09/09 Lab Meeting 心得彙整 Ingrid: (1) Preparation of GST-KR for polyclonal antibodies: protocol okay, 但把benzonase加入resuspending buffer中,而protease inhibitor放在第一次thaw完的lysate中,如果還是很黏的話要在thaw完後sonicate 15~30秒 (2)  366/367EE 有抑制現象!請再repeat一次,並且加入pFA-系列,如果也是抑制：(A) 以IP確認WT,366AA及366EE和HDAC1之interaction(we've bought this ab, didn't we?) (B) 準備做WT,366AA及366EE在293/rKSHV.219之latency disruption (3) 293/綠EBV已槓龜兩次. (4) p21/p27與老化及many others: under control! 2008/09/16 Lab Meeting 心得彙整 Ingrid:366/367EE有抑制現象, Cool. 需repeat pFA-TA系列及western（證明不是因mutant的蛋白質量表現得較少了之故).另外,可否找出先前我給妳讀的二篇paper?忘了是什麼跟Nat Cell Biol p53.謝謝.