reverse transcription PCR (RT-PCR) (Animalia)

Subcategory: gene amplification and sequencing

Papers: 1 | Mentions: 1

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Description

A multi-step molecular protocol involving RNA extraction, reverse transcription, PCR amplification using consensus primers, TA cloning, and bidirectional Sanger sequencing to characterize genetic variants in the PEPCK gene.

Typical Equipment

Ambion RNAlater
Invitrogen MiniV enzyme
Invitrogen HiFi Platinum Taq
Stratagene Robo-cyclers
Oligo 6 software
Ambion RLM-RACE kit
Qiagen kits
Qiacube processing robot
ABI BigDye 3.1 reagents
ABI 377 sequencer
BioEdit
DnaSP 5.1
TargetP server
PHASE algorithm

Output Measurements

DNA sequences
genetic variants
haplotypes

Papers Using This Protocol (1)

article

A genetic polymorphism evolving in parallel in two cell compartments and in two clades

2013BMC Evolutionary Biologyusing