Paulsson:DNA modifying enzymes

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Lurking somewhere deep within the icy confines of the -20 deg freezer at bench #2 are the following:

Contents

Restriction enzymes

AatII
Acc65I
AcuI
Afe1
AflII (Afl2)
AflIII (Afl3)
AgeI
AhdI
AscI
AsiSI
AvrII
BamHI
BamHI-HF
BclI
BglII
BlpI
BsaBI
BsiWI
BspHI
BsrGI
BssHII
BstAPI
BstXI
Bsu36I
ClaI
DpnI
EagI-HF
EcoNI
EcoRI
EcoRI-HF
EcoRV
HindIII-HF
HpaI
KpnI-HF
MfeI
MscI
NcoI
NdeI
NheI-HF
NotI-HF
NruI
NsiI
PacI
PciI
PmeI
PshAI
PstI
PvuII-HF
SacI-HF
SalI
SalI-HF
SbfI-HF
ScaI
SexAI
SgfI
SmaI
SpeI
SphI
SspI
StuI
SwaI
XbaI
XmaI
Xho1
...more information at NEB
for example: doing all your digests in Buffer #4 [1], 5 minute digestions [2], activity in Taq buffer [3], activity in Vent buffer [4], and cleavage near the end of oligos [5]

Restriction enzyme buffers

NEB: #1, #2, #3, #4, BamHI, EcoRI, 100xBSA (10mg/ml)

Polymerases

AccuPrime Pfx Supermix (Invitrogen) [6]
PCR Supermix (Invitrogen) [7]
coming in January '07... Phusion!!! not from Gilette, but from Mike Springer (include links to protein prep, protocol, plasmid map, etc...)

Other modifying enzymes

Alkaline Phosphatase, Shrimp (Roche)[8]
Mung Bean Nuclease (NEB)[9]
Plasmid-Safe DNAse [10]
DNAase from Ambion
Benzonase Nuclease from Sigma
T4 DNA Ligase [11] (and 5x buffer w/ATP) - do what you can to keep them cold at all times (eg aliquot buffer upon first use)
T4 Polynucleotide Kinase (NB: T4 PNK reaction buffer does not have ATP; add to 1mM OR use T4 Ligase rxn buf) [12]
Roche Rapid Ligation Kit [13]

DNA markers

6x TriTrack Loading Dye (Fermentas) [14]
λ DNA-Mono Cut Mix [15]
2-log ladder [16]
50 bp ladder [17]
1 Kb Plus DNA Ladder (Invitrogen) [18]
Supercoiled DNA Ladder [19]

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