Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents S U P P O R T

Protocol for Fill-in Recessed 3'-termini of Double-stranded DNA
(with T7 DNA Polymerase)

1. Prepare the following reaction mixture:
   

   10X reaction buffer	5 µl
   template DNA	2 µg
   dNTP Mix, 10 mM each	1.5 µl (0.3 mM final concentration)
   T7 DNA Polymerase	0.5 µl (5 u)
   Water, nuclease-free	to 50 µl

2. Incubate the reaction mixture at 37°C for 1.5 to 5 minutes.
3. Stop the reaction by heating at 70°C for 10 minutes.

References

1. Bodescot, M., Brison, O., T7 DNA polymerase requires unusual reaction conditions for blunt-ending activity, Anal. Biochemistry, 216, 234-235, 1994.
2. Bodescot, M., Brison, O., Efficient second-strand cDNA synthesis using T7 DNA polymerase, DNA and Cell Biology, 13, 977-985, 1994.

Ordering Information

T7 DNA Polymerase dNTP Set dNTP Mix, 10 mM each M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase RevertAid™ H Minus M-MuLV Reverse Transcriptase Nb.Bpu10I T4 DNA Ligase Water, nuclease-free

catalog@fermentas.com

Updated gegužės 09, 2007 16:40