PsyLab Meeting Report (2018/08/13) Hong CJ 6:10 pm, 2018/08/15
攜帶式睡眠血氧資料分析(fig 1, 2)：191人資料顯示，兩日居家夜眠血氧，有84%病患篩出有<90%大於5sec。+男性<90%篩出率較高85%>81%，但較嚴重個案(>20mins)，女性較高20%>16%。+隨年齡增加，較嚴重個案(>20MINS)的比例呈現上升(7%=>27%)。+診斷Sleep Apnea的篩出率為88%，Psy診斷類別篩出率78%，但Schizophrenia為93%。+Depression篩出率分別為71%及70%，Bipolar篩出率100%(N=6)。+已有Schizophrenia的Sleep Apnea研究，但尚未找到針對Sleep O2 desaturation。Further: +將無精神疾病診斷組，加入睡眠相關問題的描述並分類，進行分析。+尋找Ref後，嘗試以不同百分比下(EX:88%or92%)的缺氧時間做為標準進行分析。+目前Hypoxia和睡眠疾病的相關研究(也可從已發表的PSG研究切入)。
CRISPR: show the work flow(fig).
gRNA synthesis 有雜band，要注意annealing temperature。(這次懷疑因為PRC machine 壞掉，導致溫度不穩。)
8/1 出生的鼠寶寶總共有六隻，已經進行第二次mating (兩隻公鼠與三隻母鼠)。
gRNA Template Synthesis : got low product concentration after gel extraction. Not sure if it’s because the PCR machine breaks during PCR process. Will do gRNA template PCR again this week
Normal plasma Kynurenine level : from reference(table) , the normal plasma KYN concentration is around 500±25 ng/mL in this and many other published data; KYN, 3HAA, XA, AA plasma level might has ethnic differences in Caucasian, Hispanic, African American and Asian(notice Asian has 36% lower KYN level compare to Caucasian); African American and Asian have lower plasma 3HAA and AA levels, and lower kynureninase activity obtained from Trp-loading experiments(data not shown) suggest they might may have an innate protection mechanism involve kynureninase activity
blood genomic DNA extraction
Extract 200 μl blood sample, dissolved in 50 μl CDB.
• 200 μl blood expectation DNA weight is 8.4 μg (through the average amount of leukocytes in 200 μl blood)
Calculate the extraction rate：
Sample 1 DNA weight is 7.135 μg, extraction rate is 84.9%.
Sample 2 DNA weight is 3.308 μg, extraction rate is 39.4%.
• Extraction of the amount of DNA yield in protocol is 4-8 μg.
Sample 1 is within the yield range, Sample 2 is a little bit lower than the range.
(1) low amount of leukocytes in blood sample
(2) leukocytes amount is higher than 2 × 106 cells in 200 μl blood
(3) rupture the filter
BDNF genotyping PCR&RFLP
(1) Should notice which PCR protocol is correct.
• Lower annealing temperature will appear multiple bands.
• Wrong cycles will affect the quantity.
(2) Positive control (A/A type) should be testing at the same time. Otherwise can’t confirm enzyme digestion is completely done or not.
(3) PCR machine damage to cause wrong annealing temperature, so can see multiple bands appear on photo (slide8). It may affect RFLP’s result shows bands at 66 bps.
(1) result seems PCR didn’t be contaminated.(Blank didn’t appear any bands. )
(2) Suppose enzyme digestion is completely done, sample 1&2 are G/G type. Another two samples(郁筠&Ni) are G/A type.
(3) When pull up the comb, must be carefully to avoid breaking wells. Or bands will blurred.
Learning iPS Cells culturing Passaging
Practicing gRNA template synthesis
Preparing poster: INVESTIGATING THE ROLE OF
ZNF326 IN HUMAN NEUROBLASTOMA CELLS
PsyLab Weekly Meeting Schedule
8/20(一) 12:10pm 健凱, 琤霓, 廷偉, 亭君, Kimberly
8月lab meeting 地點