Download CG Rich Reaction Buffer (5x)

Instructions For Use / Data Sheet
IDL028
Rev Date: May 11 2015
Rev: 1
1 of 1
CG Rich Reaction Buffer (5x)
Quantity
IDL028
1 ml
Description
Templates with high-GC content are particularly difficult to amplify, due to their high
melting temperatures, and may require additional measures beyond optimizing reaction
conditions. Incomplete separation of DNA strands can adversely affect amplification
efficiency. In addition, template secondary structure or unmelted GC-rich regions can
prevent primer binding and enzymatic elongation.
Co-solvents (formamide, DMSO, and glycerol) that affect DNA melting temperature are
often used to amplify template with high-GC content, but reactions containing the cosolvents require significant optimization to prevent inhibition of Taq DNA polymerase.
GC Rich Reaction Buffer is the co-solvent for amplification of sequences that are 50% to
90% GC without inhibiting Taq DNA polymerase activity.
General Reaction
Mixture (20 l)
Template
Primer 1
Primer 2
Taq DNA Polymerase (5U/l)
10x Reaction Buffer
CG Rich Reaction Buffer (5x)
dNTP Mixture (2.5mM each)
Sterilized distilled water
1 ng-1 g
5-10 pmoles
5-10 pmoles
0.2-0.5 l
2 l
4 l
2 l
up to 20 l
Please note the above volumes are only guidelines. Optimal reaction mixture and
conditions must be determined by the end user.
Storage
≤ -18 oC
Reference
Anand, RP et al. “Overcoming natural replication barriers: differential helicase requirements”.
Nucleic Acids Research. pp. 1–15. 2011
in a constant temperature freezer.
Empire Genomics LLC 700 Michigan Ave Suite 200 Buffalo NY 14203 T 716.856.3873 F 716.856.3857 www.empiregenomics.com
For In vitro research use only.