2005-06-08

- check the filter
  --> better image if the light is not straight through
- check bernie's drawer for the filters' cases

- reading: filter, objective (numerical aperture & TIRF)

(7.50 am)
- re-install the 40x lower objective
- prepare the blank sample

(8.20 am)
- prepare the TAMRA tube --> could not see much, maybe because of photobleaching.  I accidently exposed the TAMRA tube to room light on my bench for ~3 hours

(9.21 am) take Harry's sample

Hello Rizal,

    I have prepared the samples today.  There are five tubes correspond to:

(i) ROO
(ii) SOO-H2(20)
(iii) SE-UE-VE-RE with blunt sticky ends on SE2-NTSK and RE4-NTSK
(iv) SE-UE(I1)-VE-RE with blunt ends on SE and RE
(v) SE-UE(H1)-VE-RE with blunt ends on SE and RE

I have placed them in the orange rack inside the -20C freezer.  They are in the first row.  You may need to use the tweezers (next to the gel box besides the sink) to take them out.  Please anneal them in the morning so we can use them in the afternoon.  Thanks.

Best,
Harry 

(10 am)
- anneal new samples 
  -not fluorescent 
  - TAMRA
  - FAM

2uL of 10uM SE1
2uL of 10uM SE2-DIAG
2uL of 10uM SE3-TAMRA/FAM/-
2uL of 10uM SE4-EE10
2uL of 10uM SE5
10uL of 10X TAE/Mg++
80uL of MQ H_2O

- anneal with Harry's sample

(12-5 pm)
scanning a 4 tiles tube on mica with Harry

- the 4 tiles tube with blunt ends stack with each others forming a longer tube
- it's hard to image 4 tiles tube on ROO-SOO lattice
- got a really high resolution image

(5.05 pm) 
- check the filters with 532 laser
- mount them more securely

- the lower objective was replaced with 100x not-TIRF objective
  --> it seems to me that the optical pattern decreases and the spot on the room ceiling is clearer
- still could not get any image