PsyLab Meeting Report (2018/08/06) Hong CJ 11:15 pm, 2018/08/08
CRISPR: keep amplifying. will start a new turn.
SH-SY5Y differentiation D14. 上次所提到的hetero細胞長的比KO的細胞還慢，擔心弄混了細胞株，這星期會找時間做 qPCR 或 western blot 進行確認。
山坡小屋換籠八、九月份排班：8/06-8/10: Kim & 琤霓；8/13-8/17: 健凱 & 廷瑋；8/20-8/24: 琤霓 & 亭君；8/27-8/31: 廷瑋 & Kim；9/03-9/07: 琤霓 & 亭君；9/10-9/14: 健凱 & 廷瑋；9/17-9/21: 琤霓 & 亭君；9/25-9/28: 健凱 & 廷瑋
Kynurenine ELISA(table)(本週最佳實驗): Serum Samples have matrix effect(20ng spiked Sample serum has a 36.57% recovery rate); Matrix influences results by suppression(result of 20ng spiked Diluent(2604.2ng/mL) is higher than Sample serum(1576.56ng/mL)); Sample with higher KYN conc. has stronger matrix suppression(recovery rate of 6ng and 20ng spiked Sample Serum and is 78.98% and 36.57% ); Dilution of the sample can reduce the matrix effect in this ELISA test to the acceptable range(5x Dilution Sample Serum Spiked with 6ng and 20ng has recovery rate of 92.01% and 91.88%); Still has problems with the Standard Curve or test accuracy(compare expected value of spiked Diluent with result), maybe try to compare TBS buffer with Diluent and to reduce the substrate incubation time to 25min. (Hong: 這樣寫實驗摘要非常好)
Pipetting practice: first time: CV(%) of 1000 μl to P20's 2 μl were lower than 10%, P2's 2 μl and 1 μl were both higher than 10%. Second time: CV(%) of P2's 2 μl and 1 μl were both lower than 10%.
Whole blood genomic DNA extraction: Extract 200 μl blood sample, sample 1 got DNA weight 5.708 μg, and it's extraction rate is 63.4%. sample 2 got DNA weight 3.308 μg, and it's extraction rate is 36.7%.→DNA possible fragmented while the extraction processing.(Hong: 這論點與DNA抽取率都缺乏根據)
Human BDNF PCR Gel electrophoresis:(1) gel electrophoresis result seems PCR sample wasn't contaminated.(Blank didn't appear any bands. ) (2) use longer exposure time to make photo more clear. (Hong: 這樣寫實驗摘要非常好)
Learning handling SH-SY5Y Cells and iPS Cells
PsyLab Weekly Meeting Schedule
8/13(一) 12:10pm 健凱, 琤霓, 廷偉, 亭君, Kimberly
8月lab meeting 地點