NYMU Taipei/Lab Notes/2007 10 5

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(PCR length check)
(PCR length check)
 
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== PCR length check ==
== PCR length check ==
-
* expected lengths of three ligations
+
* PCR product length without any insert = (3,189 - 3,049) + 176 = 316 by primers VF2 and VR
-
** (I) pCinRHSL (vector) + OmpRBS (insert):  
+
* expected combined insertion lengths of three ligations
-
** (II) pOmpC (vector) + TATA_INSA (insert)
+
** (I) pCinRHSL (vector) + OmpRBS (insert): 291 + 316 = 613
-
** (III) TATB_INSB (insert) + CinR+HSL+D-term (vector)
+
*** 291 = pCinRHSL (225) + Sequence after left cutting sites (1) + OmpRBS (65)
 +
** (II) pOmpC (vector) + TATA_INSA (insert): 297 + 316 = 603
 +
*** 297 = pOmpC (108) + Sequence after left cutting sites (23) + TATA (90) + INSA (66)
 +
** <s>(III) TATB_INSB (insert) + CinR+HSL+D-term (vector): 1,829 + 316 = 2,145</s>

Latest revision as of 05:17, 9 October 2007

PCR length check

  • PCR product length without any insert = (3,189 - 3,049) + 176 = 316 by primers VF2 and VR
  • expected combined insertion lengths of three ligations
    • (I) pCinRHSL (vector) + OmpRBS (insert): 291 + 316 = 613
      • 291 = pCinRHSL (225) + Sequence after left cutting sites (1) + OmpRBS (65)
    • (II) pOmpC (vector) + TATA_INSA (insert): 297 + 316 = 603
      • 297 = pOmpC (108) + Sequence after left cutting sites (23) + TATA (90) + INSA (66)
    • (III) TATB_INSB (insert) + CinR+HSL+D-term (vector): 1,829 + 316 = 2,145