Acrylamide Solutions:

-Filter all solutions with .45µm
Percentage:
6%
8%
14%
20%
23%
5X TBE:
50ml
50ml
50ml
50ml
50ml
Urea:
210.21g
210.21g
210.21g
210.21g
210.21g
40% Stock Acrylamide.:
75ml
100ml
175ml
250ml
288ml
Total Volume:
500ml
500ml
500ml
500ml
500ml
 
 
 
 
 

0.5M EDTA:

  • 10g NaOH
  • 400ml ddH2O
  • add 93g Na2EDTA while stirring
  • Increase Vol to 500ml with ddH2O
  • Check pH to 8.0, aliquot into 50ml conicals

 

1X TE:

  • 0.5ml 2M Tris (8.0) - 10mM final
  • 0.2ml 0.5M EDTA - 1mM final
  • ddH20 to final volume of 100ml. TE Low has 0.1mM EDTA.

 

5X TBE:

  • 54g Tris Base
  • 25.5g Boric Acid
  • 20ml 0.5M EDTA (8.0)
  • ddH2O to final volume of 1L. (pH 8.3)

 

50X TAE:

  • 242g Tris Base
  • 57.1ml Glacial Acetic Acid
  • 100ml 0.5M EDTA (pH 8.0)
  • water to 1L

 

FLB: (Formamide Loading Buffer)

  • 4.75ml 95% Formamide
  • 150µL 0.5M EDTA
  • 50µL 2.5% Bromophenol Blue
  • 50µL 2.5% X.C.
  • Make 100µL aliquots, filter thru large syringe, 0.2µm.

 

Reverse Transcriptase (RT) Buffer:

25mM KCl 83µL 3M KCl
50mM Tris (7.5) 250µL 2M Tris
10mM DTT 100µL 1.0M DTT
3.5mM MgCl2 35µL MgCl2
0.5mM all 4 dNTPs 50µL each dNTP @ 100mM
100µg/mL BSA 50µL 20mg/mL BSA
ddH2O to 10mL
  • Filter, make ~600µL aliquots