Noi/NOTES/2012-7-31: Difference between revisions

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>Noi
(Created page with "http://genome-tech.ucsd.edu/LabNotes/index.php/noi:DMR220k_LabNotes '''Link to calendar'''<br> = Preparation of sheared gDNA for N37 sample set = * Alice make an appointme...")
 
>Noi
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* I need to check if there is some evaporation and need to bring total volume to 8ul with EB buffer.
* I need to check if there is some evaporation and need to bring total volume to 8ul with EB buffer.
* I plan to shear 200ng of gDNA, so I will take 2ul of gDNA and add to 48ul of H2O.
* I plan to shear 200ng of gDNA, so I will take 2ul of gDNA and add to 48ul of H2O.
* The total volume reduced to ~5ul, I added 3ul to each tube to bring volume up to 8ul
{| {{table}} border = 1
{| {{table}} border = 1
| align="center" style="background:#f0f0f0;"|'''Sample IDs'''
| align="center" style="background:#f0f0f0;"|'''Sample IDs'''

Revision as of 01:07, 31 July 2012

[Link to calendar]

Preparation of sheared gDNA for N37 sample set

  • Alice make an appointment with James at Biogen

Sample list and code on the tube

  • From the previous experiment, gDNA of the 10 tissues of N37 were diluted to 100ng/ul total volume 20ul. I used 12 ul for BSPP assays, and have 8ul left.
  • I need to check if there is some evaporation and need to bring total volume to 8ul with EB buffer.
  • I plan to shear 200ng of gDNA, so I will take 2ul of gDNA and add to 48ul of H2O.
  • The total volume reduced to ~5ul, I added 3ul to each tube to bring volume up to 8ul
Sample IDs Sample code on tubes Original conc. (ng/ul) Volume
for 200ng (ul)
EB (ul) Total volume (ul)
N37-Cerebellum (CRBL) N37-1 ~100 2.00 48.00 50.00
N37-Colon N37-2 ~100 2.00 48.00 50.00
N37-Frontal lobe (FL) N37-3 ~100 2.00 48.00 50.00
N37-Heart N37-4 ~100 2.00 48.00 50.00
N37-Small intestine (SI) N37-5 ~100 2.00 48.00 50.00
N37-Liver N37-6 ~100 2.00 48.00 50.00
N37-Lung N37-7 ~100 2.00 48.00 50.00
N37-Skeletal muscle (SM) N37-8 ~100 2.00 48.00 50.00
N37Pancrease N37-9 ~100 2.00 48.00 50.00
N37Stomach N37-10 ~100 2.00 48.00 50.00
 Follow up the set up for Covaris shearing (From BioGem and Alan's note: [[1]]
Target BP 400
Tube Micro
Duty Factor 10%
Peak Incident Power (w) 140
Cycle per Burst 200
Time (secs) 55
Volume 50ul
Temperature (c) 7 
Intensifier Yes