Sequencing Reaction Clean-Up Kit

Overview

Purify sequencing extension products from dye terminators, primers and other contaminants
Also purify DNA from different enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations.
High recovery
Fast and efficient spin column format
Also available in 96 well format for high throughput

This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.

The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up. Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.

Details

Supporting Data

Figure 1. Sequencing Chromatogram of pDC-CG DNA. A single extension product was divided into 3 equal aliquots and was cleaned using either Norgen's Sequencing Reaction Clean-Up Kit, or one of two competitor kits. Sequencing data was then generated using Bigdye Terminator chemistry on the Applied Biosystems 3130xL DNA Sequencer. Panel A corresponds to the results when the sample was cleaned using Competitor 1, and this chromatogram indicates low extension product yields. Panel B corresponds to the results when the sample was cleaned using Competitor 2, and this product exhibited poor dye terminator removal that introduced several blobs to the sequence. Panel C corresponds to the sample cleaned with Norgen’s Sequencing Reaction Clean-Up Kit, and shows good yield and efficient removal of dye terminator and contaminants as indicated from the chromatogram signals.

Kit Specifications - Spin Column
Column Binding Capacity	13 μg
Maximum Column Loading Volume	650 μL
Maximum Amount of Starting Material	25 μg
Time to Complete 10 Purifications	10 minutes
Minimum Elution Volume	30 μL

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 34500 (50 preps)	Cat. 34400 (2 x 96-well plate)
Binding Buffer C	30 mL	3 x 30 mL
Wash Solution A	12 mL	2 x 20 mL
Elution Buffer B	8 mL	2 x 15 mL
Spin Columns	50	-
Collection Tubes	50	-
Elution Tubes (1.7 mL)	50	-
96-Well Plate	-	2
Adhesive Tape	-	4
96-Well Collection Plate	-	2
96-Well Elution Plate	-	2
Product Insert	1	1

Documentation

PROTOCOLS

(34400) Sequencing Reaction Clean-Up 96-Well Kit (High Throughput) - Protocol (96-well)

(34500) Sequencing Reaction Clean-Up Kit (Spin Column) - Protocol (50 prep)

SAFETY DATA SHEETS

34500 - Sequencing Reaction Clean-Up Kit

34400 - Sequencing Reaction Clean-Up 96-Well Kit