Team:Heidelberg/AwesomeSheet/Enzymes

From 2014.igem.org

– Enzymes

ABCDEFGHIJKLMNOPQRSTUVW
1
IDName
Concentration
BufferInkubation Inaktivation
max DNA conc.
CommentBSAATPUnit Defnition
2
correct useXNameXTtTt
3
c/(u/µL)
/((U/uL)/(ng/uL))
°Cmin°Cminc/(ng/ul)1.12.23.1
CutSmart
T7 Lig Buffer
4
EEcoRI-HF200.04371565202500%100%0%100%One unit is defined as the amount of enzyme required to digest 1 μg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 μl.
5
XXbaI200.04371565202500%100%75%100%One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam-/HindIII digest) in 1 hour at 37°C in a total reaction volume of 50 µl.
6
SSpeI-HF100.023715652025
nur 5 x overdigestion
075%100%25%100%One unit is defined as the amount of enzyme required to digest 1 µg of Adenovirus-2 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
7
PPstI-HF200.043715652025010%75%50%100%One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam-/HindIII digest) in 1 hour at 37°C in a total reaction volume of 50 µl.
8
T4LT4 DNA Ligase4002
10xT4LBuffer
1016106510101
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5´ DNA termini concentration of 0.12 µM, 300- µg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
The unit definition uses 0.12 μM (300 μg/ml) lambda HindIII fragments. The high DNA concentration can be used for linker ligation. To promote circle formation, useful in transformation, a lower total DNA concentration should be used. The overall concentration of vector + insert should be between 1-10 μg/ml for efficient ligation. Vector:Insert molar ratios between 1:1 and 1:10 are recommended (1:3 is typical). If you are unsure of your DNA concentrations, perform multiple ligations with varying ratios.
9
T4LHCT4 DNA Ligase h.c.2000216106510101
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5´ DNA termini concentration of 0.12 µM, 300- µg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA Ligase Reaction Buffer.
300
10
T7LT7 DNA Ligase300015
2xT7LBuffer
22530donot10
needs purification before electroporation
11One unit is defined as the amount of enzyme required to give 50% ligation of 100 ng HindIII fragments of λ DNA in a total reaction volume of 20 μl in 30 minutes at 25°C in 1X T7 DNA Ligase Reaction Buffer.5
11
T7LvT7 DNA Ligase400152530donot10
needs purification before electroporation
11One unit is defined as the amount of enzyme required to give 50% ligation of 100 ng HindIII fragments of λ DNA in a total reaction volume of 20 μl in 30 minutes at 25°C in 1X T7 DNA Ligase Reaction Buffer.
12
Flex MM
Phusion Flex Mastermix
2nonoOne unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluBIe material in 30 minutes at 74°C
13
OneTaqOneTaq Master Mix2
never
ever
14
Flash MM
Phusion Flash Master Mix
2
15
KKpnI-HF200.043715
not in your
wildest dreams
25100%25%5%100%
16
BsaIBsaI100.0437606520.00175%75%100%100%
17
BsmBIBsmBI-HF100.045515802025010%50%100%25%One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 55°C in a total reaction volume of 50 µl.
18
BglIIBglII100.0437nope10%10%100%10%
19
XmaIXmaI100.043715652025%50%10%100%One unit is defined as the amount of enzyme required to digest 1 µg adenovirus-2 in 1 hour at 37°C in a total reaction volume of 50 µl.
20
BsaI-HFBsaI-HF200.0437156520150%100%25%100%