---
author: Sam White
toc-title: Contents
toc-depth: 5
toc-location: left
date: 2011-04-11 21:35:32+00:00
layout: post
slug: ligations-cox1cox2-pcr-products
title: Ligations - COX1/COX2 PCR Products
categories:
  - 2011
  - Miscellaneous
tags:
  - COX
  - COX1
  - Crassostrea gigas
  - cyclooxygenase
  - ligation
  - Pacific oyster
  - pCR2.1
  - PGS
  - PGS1
  - prostaglandin synthase
  - TOPO TA Cloning
---

Performed ligations/cloning on a variety of COX1 genomic and COX 5' RACE products using the TOPO TA Cloning Kit (Invitrogen). Used 2uL of gel-purified PCR product in each cloning rxn, 2.5uL of H2O, 1uL of salt solution, and 0.5uL of the Invitrogen pCR2.1 vector. Incubated samples for 5mins at RT. Used 2uL of the cloning reaction to transform TOP10 chemically competent cells (Invitrogen), mixed very gently, incubated on ice for 5mins, heat shocked at 42C for 30s and immediately placed on ice. Added 250uL of SOC Medium and incubated tubes at 37C, 200RPM for 1hr. Plated 50uL of each transformation on LB+Kan plates (made by Steven on unknown date with unknown Kan concentration) containing 40uL of 40mg/mL X-gal. Incubated O/N at 37C. Remaining volume of transformed bacteria were stored @ 4C.
