Difference between revisions of "Mc4r-Cone Genotyping"

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!| PCR Step || Temperature || Time || Cycles || Notes
 
!| PCR Step || Temperature || Time || Cycles || Notes
 
|-
 
|-
1. Initiation/Melting || 94 || 5:00 || 1 ||
+
|1. Initiation/Melting || 94 || 5:00 || 1 ||
 
|-
 
|-
2. Denaturation || 94 || 0:45 || 40 ||
+
|2. Denaturation || 94 || 0:45 || 40 ||
 
|-
 
|-
3. Annealing || 66 || 0:45 || 40 ||
+
|3. Annealing || 66 || 0:45 || 40 ||
 
|-
 
|-
4. Elongation || 72 || 0:45 || 40 || steps 2-4 to be run in sequence
+
|4. Elongation || 72 || 0:45 || 40 || steps 2-4 to be run in sequence
 
|-
 
|-
5. Amplification || 72 || 7:00 || 1 ||
+
|5. Amplification || 72 || 7:00 || 1 ||
 
|-
 
|-
6. Finish || 4 || infinite || n/a ||
+
|6. Finish || 4 || infinite || n/a ||
 
|}
 
|}
  
 +
More information here [https://drive.google.com/file/d/0B1nvEJ5tMFVtdnJrSG91aFNhbHRjaUh0RGxLa1lUZ2tXWDhn/view mc4r-Cone]
 
[[ Category:Genotyping ]]
 
[[ Category:Genotyping ]]
 
[[ Category:Mouse Work]]
 
[[ Category:Mouse Work]]

Latest revision as of 17:54, 5 June 2020

PRIMERS

  • Fwd: CAC TCG GAG CTT CCC TGA CCC AG
  • Rev: GAC CAT GGT TTC CGA CCC ATT
  • Neo Rev: TTC CAA GCC TCT GAG CCC AGA

Make primers at 1uM (Fwd + Rev and Fwd + Neo Rev). PCR primers separately.

PCR PROTOCOL

PCR Step Temperature Time Cycles Notes
1. Initiation/Melting 94 5:00 1
2. Denaturation 94 0:45 40
3. Annealing 66 0:45 40
4. Elongation 72 0:45 40 steps 2-4 to be run in sequence
5. Amplification 72 7:00 1
6. Finish 4 infinite n/a

More information here mc4r-Cone