Hosuk:LabNotes/2013-11-21: Difference between revisions

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(Created page with "*LabNote ===Padlock Probe Concentration Test=== *Sample : 96 well plates, made at 10/23 *2048 x 2048, 600Hz *Each stripping : 30min, and annealing : 15m...")
 
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{| {{table}} border = 0
{| {{table}} border = 1
| align="center" |'''Pos 6, 5uM'''
| align="center" |'''Pos 6, 5uM'''
| align="center" |'''Pos 1, 5uM'''
| align="center" |'''Pos 1, 5uM'''
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|-
|-
| align="center" | 1st Rolony || align="center" | 7387 || align="center" | 6495 || align="center" | 6040 || align="center" | 6089 || align="center" | 7054 || align="center" | 8663
| align="center" | 1st Rolony || align="center" | 7387 || align="center" | 6495 || align="center" | 6040 || align="center" | 6089 || align="center" | 7054 || align="center" | 8663
|-
| align="center" | MALAT1 || align="center" | 4886 || align="center" | 4196 || align="center" | 3817 || align="center" | 4155 || align="center" | 6179 || align="center" | 5561
| align="center" | MALAT1 || align="center" | 4886 || align="center" | 4196 || align="center" | 3817 || align="center" | 4155 || align="center" | 6179 || align="center" | 5561
|}
|}

Revision as of 22:55, 21 November 2013


Padlock Probe Concentration Test

  • Sample : 96 well plates, made at 10/23
  • 2048 x 2048, 600Hz
  • Each stripping : 30min, and annealing : 15min
  • The concentration of the dye-labeled probe : 1uM
  • Use “Mark and Fine” function to save each location at each well


Procedure

  • Process Steps
    1. Step 1 : Image 1st Rolony
    2. Step 2 : Strip, anneal ppMATLA1 (0.1nM, 1nM, 10nM, 100nM, and 5uM)
    3. Step 3 : Add “Shifted FISSEQ_2ndRCAprimerDye” , image


Result

  • Need overlap counting, but
  • 0.1nM of padlock probe seemed many signal, and Need to plat toe…
  • Will do more further analysis


1st Rolony Count
  • Pos 6 : for strip test


Pos 6, 5uM Pos 1, 5uM Pos 2, 100nM
Pos 5, 10nM Pos 4, 1nM Pos 3, 0.1nM


Counting
Dye Probes Pos3 Pos4 Pos5 Pos2 Pos1 Pos6
0.1nM 1nM 10nM 100nM 5uM 5uM
1st Rolony 7387 6495 6040 6089 7054 8663
MALAT1 4886 4196 3817 4155 6179 5561