13A BioSAXS

蛋白質樣品相關Q&A     BioSAXS Q&A

Q1.樣品的濃度與體積需求?    Sample requirements?

Ans : SEC-SAXS實驗,假設分子量Y (kDa),原則上建議需要濃度500/Y (mg/mL)以上。(舉例: BSA 66.8 kDa,則需至少7.5 mg/mL) 體積方面,初步測試SEC-SAXS只需10 µL,完整收SEC+SAXS時一次的注射量是50 -100 µL。                                                                                                                                                                                                                       

Ans : For the SEC-SAXS experiment, say your protein has a MW of Y kDa; we'll suggest the concentration to be at least (500/Y) mg/mL. (For example, BSA monomer is 66.8 kDa, then you'll need at least 7.5 mg/mL to get a good S/N ratio) A pre-run of SEC-SAXS requires only 10 µL, and an entire production run needs about 50-100 µL.                                                                                                                                                                 

Q2.除了樣品外要攜帶的物品及本實驗站還有提供哪些實驗器材?    Any other items I need to bring to the beamline?

Ans:請記得帶緩衝液,一個樣品搭配的緩衝液請帶至少250 mL以防萬一。帶來前請先用0.22 um filter過濾過。實驗站及T260實驗室備有pipette、離心機、4度冰箱、超音波震盪槽、pipette tips、Eppendorf tubes、HPLC進樣用的insert與樣品瓶、離心過濾管、濃縮管、純水機、天秤等基本設備,如有特殊需求也可以直接跟我們聯絡。                                                                                                                                     

Ans : Please bring the suitable buffer solution for your protein, 250 mL for each sample, filtered by 0.22 um filter. 13A endstation and T260 lab have pipette, pipette tips, centrifuge, freezer, sonicator, Eppendorf tubes, HPLC sample vials, centrifugal filters, concentrator tubes, Millipore-water, analytical balance, etc. Contact us if you have special needs.                                                                                           

 

Q3.可以安裝的管柱種類?    What column choice do I have?

Ans:如下圖。    Here is a list of the columns we can install to our system:   

 

                           Agilent SEC-03            Tosoh-PWxl      

 Superdex-75         Superdex-200                         Wyatt   

材料樣品相關Q&A     Material science-related sample Q&A

Q1.可以使用的樣品載台與控溫範圍?    What types of sample holders (cells) do you provide? How about the temperature tuning region?

Ans: 如下圖。    Here are some pictures of the cells you can use with their temperature tuning range:

Liquid cell: 80 µL

Capillary cell: 2 mm i.d. needs at least 80 µL

Linkam Temperature Control Stage: -195 to 600 °C

 

Q2.可達到的最大q值與最小q值?     What are the qmax and qmin ?

Ans:

Highest qqmax = 3.5 Å‑1

Lowest qqmin = 0.0004 Å-1

注意:單次量測無法涵蓋這麼廣的區間   

Note: A single measurement can't cover qmin to qmax. It's too broad.                                                                                                                       

SAXS-WAXS 組合的單次 動態q-range (qmax/qmin ~250)

There is a dynamic q-range (qmax/qmin ~250) for one measurement.                                                                                           

例如 : 選定qmin = 0.01 Å-1; qmax = 2.5 Å-1 

          選定qmin = 0.001 Å-1; qmax = 0.25 Å-1 

For example,  if you've selected qmin = 0.01 Å-1; then qmax = 2.5 Å-1 in one shot.

If you choose qmin = 0.001 Å-1; qmax = 0.25 Å-1.                                                                                            

如何利用蛋白質分子量M (in kDa)估算欲量測尺度的 q-range:

(How to estimate the q-range for a protein with the MW of M kDa?)                                                                                                                                                                                            

Rg (nm)= 10^{0.42*[log M]-0.32}

qmin ~ 1/(2Rg)

For example : 67 kDa (BSA);  Rg = 2.79 nm ; qmin = ~ 0.018 Å-1.

 J.Appl. Cryst. (2015). 48, 1604–1606

共同Q&A     General Q&A

Q1.實驗結束後若有發現忘記複製的數據要去哪邊抓取或是重新做data reduction?     Where can I download my data or re-do the data reduction?

Ans:Please go to the DiCOS-BioSAXS Platform; here is the link: https://bioswan.twgrid.org/

Tutorial video : https://youtu.be/l3eat9rUlxI

 

Q2.實驗過程中出現問題該如何聯繫實驗站負責人員?     How do I contact endstation people if there are any problems during my beamtime?

Ans:請利用我們的視訊系統送出求救訊號。    Please use our webcam system to request assistance.

教學影片 : (待上)

 

Q3.實驗結束後用戶應該關閉哪些儀器?     What do I have to do after my experiments (before leaving NSRRC)?

Ans:Please follow the steps shown in this tutorial video : https://youtu.be/c8QTWeoX640

 

Q4.離實驗站最近的出入大門?     What's the closest gate number to TPS13A?

Ans:23號門,訂購外送餐點或上下大型貨物可以利用此門。(疫情期間外送餐點只能送到大門警衛處)

Ans:Gate #23, you can ask the food delivery driver to drop stuff there or load oversized cargo through that gate.

(However, the food can only be delivered to the main gate if the pandemic situation gets worse.)

當發生緊急事故必須進行疏散時,請大家就近前往下圖的紅色1號疏散集合點,以利人員清點與後續應變措施。

If there is any emergency and an evacuation is needed, please go to the red spot (1) shown below for further instructions.

 

Q5.緊急聯繫或發生各種況狀可能會用到的分機號碼?     What are the extension numbers I might need to know?

Ans:警衛室 Ext. 3333 (計程車/招待所鑰匙/緊急事故)

          醫務室 Ext. 8217

          TPS控制室 Ext. 8201

          Dr.葉奕琪 Ext. 7106

          Dr.施怡之 Ext. 7156

          TPS'13A光束線 Ext. 2132

          T262A休息室 Ext. 8262 備有咖啡機/飲料/點心,實驗站有氣泡水機。

          T260生物樣品實驗室 Ext. 8260 (需事先申請)

          T244A材料樣品準備室 (此實驗室專門處理有機溶劑請多加利用,需事先申請)                                                                                            

Ans:Security office Ext. 3333 (For calling taxi/guest house key deposit/emergency)

          Medical office Ext. 8217

          TPS control room Ext. 8201

          Dr.Yi-Qi Yeh Ext. 7106

          Dr.Orion Shih Ext. 7156

          TPS'13A endstation Ext. 2132

          T262A break room Ext. 8262

          (we have a coffee machine/beverages/snacks there for free, the endstation has a soda maker for sparkling water lovers)

          T260 Bio-lab Ext. 8260 (BSL-2 lab, please apply before use)

          T244A lab (General lab, you can handle organic solvents there, but also need pre-apply for usage)

 

 

 

 

 

 

 

 

 

 

 

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Endstation

+886-3-578-0281 Ext. 2131

BL Spokesperson

Dr. U-Ser, Jeng (鄭有舜)
usjeng@nsrrc.org.tw
Ext. 7108

BL Local Contact/Manger

Dr. Yi-Qi, Yeh (葉奕琪)
yeh.yq@nsrrc.org.tw
Ext. 7106


Techniques

  • Small Angle X-ray Scattering (SAXS)

Disciplines

  • Structural biology
  • Polymer and soft matter

General Information

  • Source: 13A
  • Energy Range: 5.6-25 keV for DCM
  • Focused Spot Size: 400 x 200 μm²
  • Status: under construction







National Synchrotron Radiation Research Center

101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu, Taiwan 30076

+886-3-578-0281