Haynes Lab:Notebook/Jan/2015/04/06: Difference between revisions

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----
'''Sequencing: BD006/MV2'''


 
{|
 
|-
| Sample # || Template || Primer || Benchling ref file
|-
| 1. || BD006MV2_1 || DD122B || [https://benchling.com/s/MRaqMtbX/edit ChromatinSensor1_MV2]
|-
| 2. || BD006MV2_1 || DD123B || "
|-
| 3. || BD006MV2_2 || DD122B || "
|-
| 4. || BD006MV2_2 || DD123B || "
|-
| 5. || BD006MV2_3 || DD122B || "
|-
| 6. || BD006MV2_3 || DD123B || "
|-
| 7. || BD006MV2_4 || DD122B || "
|-
| 8. || BD006MV2_4 || DD123B || "
|}




Revision as of 11:18, 6 April 2015

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04/06/15

  • Assembly - DPRE into "BD004"
  • Sequencing - BD006/MV2


Assembly


  • Calculations
    • Vector - delBD004(E/X); 50 ng = 7.7 μL
    • Insert - DPRE(E/S); (1740/7337)*2*50 = 23.7 ng = 1.4 μL


  • Ligations
  1. DPRE + delBD004
  2. delBD004 only (neg)


Reagent 1 2
DNA insert 1.4 ---
DNA vector 7.7 1.5
2x buffer 9.6 9.6
T4 ligase 0.5 0.5
dH2O --- 1.4
  19.2 μL 19.2 μL

--> RT/ ~30 min.

  • Add 50 μL DH5-α turbo
  • 5 min./ ice
  • Plate on 100 ug/mL amp agar. Grow at 37 °C o/n


RESULTS (04/06/15)

  1. tba


Sequencing: BD006/MV2

Sample # Template Primer Benchling ref file
1. BD006MV2_1 DD122B ChromatinSensor1_MV2
2. BD006MV2_1 DD123B "
3. BD006MV2_2 DD122B "
4. BD006MV2_2 DD123B "
5. BD006MV2_3 DD122B "
6. BD006MV2_3 DD123B "
7. BD006MV2_4 DD122B "
8. BD006MV2_4 DD123B "



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