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Which are the cheap restriction enzymes?


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5 replies to this topic

#1 seanspotatobusiness

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Posted 07 August 2009 - 12:55 AM

I'm new to molecular cloning and I'd like a list that I can import into Vector NTI or Clone Manager that consists of just the relatively cheap restriction enzymes, so I can use those for analysis, where possible. Does anyone have such a list already? I'm pretty new to the programs, but I think there's some way I could import the list and then look for those sites within my molecules. Thanks! :P

#2 phage434

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Posted 07 August 2009 - 03:19 AM

You want not just cheap but good (the manufacturers will tell you they are all good, but they are not).

Here's a short list:
EcoRI, PstI, BamHI, BglII, HindIII, XhoI
I'd recommend the -HF versions of these enzymes from NEB.

People have trouble with SalI.

XbaI and SpeI are reliable but relatively expensive.

#3 Andriy

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Posted 07 August 2009 - 06:36 AM

Cheap enzymes are the ones you express and purify in your lab :-)

Of course, if you live in a country where patents are not an issue

#4 seanspotatobusiness

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Posted 07 August 2009 - 08:01 AM

Cheap enzymes are the ones you express and purify in your lab :-)

Of course, if you live in a country where patents are not an issue


Where do you live Andriy? :)

#5 noelmathur

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Posted 07 August 2009 - 11:37 PM

I use EnzymeX on Mac so I am not familiar with your programs but IMO, you are starting in wrong fashion. You should not make list of 'cheap' enzymes. You should make a list of enzymes those you need. We buy enzymes as we need and our current availability is as follows, mostly Fermentas but few NEB and Takara.

AluI
AgeI (BshTI)
BplI
BsaHI
BclI
BamHI
BglII
BcuI
BfiI
Bsp143II
Bsu36I
CfrI
Cfr9I
ClaI
Cfr42I
Eco32I
EcoRI
Eco 91I
Eco52I
Eco88I
Eco47III
Eco81I
HindIII
Hin1I
HincII
KpnI
LguI (SapI)
MluI
NcoI
NheI
NotI
NsiI
NgoMIV
PaeI
PstI
PvuI
PciI
PvuII
SmiI
SmlI
SmaI
SacI
ScaI
VspI
XbaI
XhoI
XcmI

Out of these, I avoid Cfr9I, its tricky to use.
SphI (PaeI) is very expensive enzyme and I avoid using that one as well because once my boss came to me showing how much he is paying for PaeI. I got super pissed as I never waste resources, yet he comes to me showing the bill :huh: (and others were using the same tube and he shows it to only me)

#6 seanspotatobusiness

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Posted 09 August 2009 - 05:32 AM

I use EnzymeX on Mac so I am not familiar with your programs but IMO, you are starting in wrong fashion. You should not make list of 'cheap' enzymes. You should make a list of enzymes those you need. We buy enzymes as we need and our current availability is as follows, mostly Fermentas but few NEB and Takara.

Out of these, I avoid Cfr9I, its tricky to use.
SphI (PaeI) is very expensive enzyme and I avoid using that one as well because once my boss came to me showing how much he is paying for PaeI. I got super pissed as I never waste resources, yet he comes to me showing the bill :unsure: (and others were using the same tube and he shows it to only me)


I guess you misunderstand my intentions. When I want to do a restriction analysis to determine whether I have an insert in the correct orientation for example, I'd like to be able to tell my computer program to search for sites which can be cheaply cut. If I cannot find any suitable sites for cutting, then I can open the gates to the more expensive enzymes.




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