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Description
The Internal Lane Standard 600 marker is a stock solution consisting of 22 DNA fragments
ranging in size from 60–600bp. Size fragments from 60–200bp are spaced at 20bp intervals,
while the fragments from 200–500bp and from 500–600bp are spaced every 25 bases and 50
bases, respectively. Fragments that are multiples of 100 bases are present in approximately
twice the yield of other fragments to simplify size assignment. The DNA ladder is double
stranded and asymmetrically labeled with carboxy-x-rhodamine (CXR).
The Internal Lane Standard 600 marker is intended to be used in assigning sizes to DNA
fragments separated by electrophoresis and detected using a variety of fluorescence-detecting
instruments (e.g., the Hitachi FMBIO® Fluorescent Scanner and the ABI PRISM®
310 Genetic Analyzer). The marker comes with Blue Dextran Loading Solution and Bromophenol
Blue Loading Solution.
An electrophoretic profile of the ILS 600 detected with an ABI PRISM® 310 Genetic Analyzer
is shown in Figure 1. An image of this marker on a 5% denaturing Long Ranger™ polyacrylamide
gel detected with the Hitachi FMBIO® II Fluorescence Imaging System is shown in Figure 2.
Features
Superior Performance:- Comparisons by Dr. Ryszard Pawlowski of internal lane standards from Promega and Perkin-Elmer
Corporations have shown that Promega's standards provide unsurpassed consistency in size
determination of Profiler Plus™ alleles. A comparison of size determination of those alleles
by Promega's Fluorescent Ladder (CXR), 60–400 Bases (which contains a subset of the fragments
in the ILS 600), and the Genescan® 500 [ROX] internal lane standard shows that Promega's
internal lane standard has significantly less deviation in fragment size assignment as
shown in Figure 3. This work was performed using an ABI PRISM® 310 Genetic Analyzer.
In the case of the Promega Fluorescent Ladder (CXR), 60–400 Bases, the size deviation
varied by no more than 0.2bp across all allele sizes. In the case of the Genescan® 500,
the size deviations at small alleles were twice—and at larger alleles were five times—
those determined using the Promega size ladder. In some cases the average size deviation
using the Genescan® 500 ladder was nearly 1bp.
The ILS 600 takes the superior performance of Promega's internal lane standards to a new
level: Because the ILS 600 contains all of the bands found in the Fluorescent Ladder (CXR),
60–400 Bases, plus an extended size range up to 60 bases, this consistency in size assignment
is available over a greater range of fragment sizes.
Increased Precision of Analysis:- Use of this marker reduces the number of allelic ladder lanes needed per gel
and, therefore, increases the number of lanes available for amplified sample materials.
Separate Detection:-
Because each fragment is labeled with CXR, the ILS 600 may be detected separately
(as a third color) in the presence of fragments labeled with fluorescein and
carboxy-tetramethyl-rhodamine (TMR) using the Hitachi FMBIO® and
FMBIO® II Fluorescence Imaging Systems.
Simple Interpretation:-
Because fragments that are multiples of 100 bases have approximately
twice fluorescence of other fragments, size assignment is simplified.
System Components
- » Internal Lane Standard 600 (150μl)
- » Blue Dextran Loading Solution (1ml)
- » Bromophenol Blue Loading Solution (1ml)
FAQ
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Related Information
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