Difference between revisions of "Mc4r-Cone Genotyping"

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Revision as of 17:48, 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

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 ||

PCR Step	Temperature	Time	Cycles	Notes