OPRM1_pGHE Subcloning (Zinc)

Include the date and initials of who did each part of the procedure. Keep good documentation with images and notes with each step.

Plasmid

Insert description of plasmid and pictures of insert and vector.

Primers

HiFi Primers

OPRM1_HiFi_FOR

OPRM1_HiFi_REV

pGHE_OPRM1_HiFi_FOR

pGHE_OPRM1_HiFi_REV

Name, Sequence, Length, GC%, Tm, and where they were purchased. Include images of primers in plasmid.

PCR

Notes of what occurred during the protocol, the protocol, and what was included in each mixture.

Dpn1 Digestion

Protocol, concentration, and length digested.

HiFi Reaction

Protocol, notes, what was in each mixture and how long was it ran.

Gel Electrophoresis to Confirm in Plasmid

Electrophoresis protocol and any adjustments made, what went into each tube, image of gel, and analysis of results. Include table as well.

Transformation

Protocol for regular and negative control. Maybe a positive control too? Images and notes.

Inoculation

Protocol and exact time spun. Images and notes.

TENS Prep and Ethanol Precipitation (MaxiPrep)

Protocol, notes, images, and any adaptations.

Gel Electrophoresis to Determine Concentration

Protocol, tube mixture, analysis, gel image and table.

Sequencing

Protocol, services utilized, images of results, and concentrations submitted.

Transcription

Protocol, images, and notes