Korea U Seoul/1 October 2010

From 2010.igem.org

(Difference between revisions)
(New page: ===Digestion=== <br><BR> # Check : recombinant plasmid sequence # Selection of correct clones # Digestion of PCR products(ParsR-GFP) and pSB1C3 ## ParsR-GFP : ''Eco''RI and SpeI ## pSB1C3 ...)
 
Line 26: Line 26:
|12.5 / total = 50
|12.5 / total = 50
|-
|-
-
!  colspan="2"     Completely digestion at 37˚C for 2 hours (at least)
+
!  colspan="2"   |  Completely digestion at 37˚C for 2 hours (at least)
<br>    and stop at 80˚C for 20min     
<br>    and stop at 80˚C for 20min     
|}<br>
|}<br>

Latest revision as of 13:42, 20 October 2010

Digestion



  1. Check : recombinant plasmid sequence
  2. Selection of correct clones
  3. Digestion of PCR products(ParsR-GFP) and pSB1C3
    1. ParsR-GFP : EcoRI and SpeI
    2. pSB1C3 : EcoRI and PstI


Reagent Volume (μL)
DNA (about 30ng/μL) 30
10x NEB buffer 2 5
BSA (10mg/mL) 0.5
Appropriate 1st and 2nd restriction enzymes 2 (each 1)
D.W. 12.5 / total = 50
Completely digestion at 37˚C for 2 hours (at least)


and stop at 80˚C for 20min


  1. Confirmation of digested products by agarose gel electrophoresis (Figure 6)
  2. Quantification of DNA concentration by NanoDrop
Retrieved from "http://2010.igem.org/Korea_U_Seoul/1_October_2010"