---
title: Sanger Sequencing Quick Tips Guide | GENEWIZ from Azenta Life Sciences
description: Tips and tricks for troubleshooting a poor sequencing reaction from a PCR template, producing and purifying a single-band PCR product.
---

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# Sanger Sequencing Quick Tips Guide

QUICK TIPS GUIDE

# Sanger Sequencing Quick Tips Guide

Tips and tricks for troubleshooting a poor sequencing reaction from a PCR template:

#### Failed reactions (i.e., no priming)

Usually indicated by a chromatogram with a signal intensity of 100.

#### Nonspecific reactions

Characterized by two or more overlapping traces in the chromatogram that represent different populations of sequencing products.

#### Spectral pull-up

Occurs when the signal intensity of the sequencing reaction is so strong that the fluorescence signal effectively spills over into the other collection channels.

![Untitled design (58)](https://web.genewiz.com/hs-fs/hubfs/Untitled%20design%20(58).png?width=2000&height=2000&name=Untitled%20design%20(58).png "Untitled design (58)")

## Download Our Guide

## The Sanger Quick Tips Series Also Includes:

#### Volume 1: *Producing a Single-band PCR Product*

- PCR primer design tips and guideline
- Polymerase enzyme selection pros and cons
- Buffer and cycling parameters
- PCR adjunct selection

#### Volume 3: *Purifying a Non-specific PCR Product*

- Isolation of a single-band by gel purification
- Single-band recovery using band-stab PCR
- Purifying a single product using nested primers

#### Volume 2: *Purifying a Single-band PCR Product*

- Enzymatic PCR clean-up
- Purification by DNA-binding matrix

#### Volume 4: *PCR Visualization by Gel Electrophoresis*

- Running buffer selection
- Voltage and run-time optimization
- Loading dyes
- Target agarose percentages
- Troubleshooting common gel problems