This is the latest MolBi list of restriction enzymes for ApE. It contains only the enzymes available through Thermoscientific (including the FastDigest enzymes having new names) and NEB. It also contains the latest MolBi REs group.

About our home-made Taq polymerase ("LunaTaq"):
it is now 3 years old, and appears to become less active.
Advice: do not use 0.2 -0.5 u per PCR reaction, but use 1 to 1.5 u instead...

October, 20   2011 NEWS

A major update of the MolBi  REnzymes page is done.
The list of all commercially available REs, showing which of them are present at MolBi, is now made from the Restriction Enzymes pages from the Fermentas catalogue.
Prices of the FD enzymes can be a guideline for you, when you are free to choose REs for example to screen plasmids from bacterial transformants.
For Gateway transformants, StyI may be a useful and cheap RE. See the list with the REs sorted by price!

August, 23   2011 NEWS

A printable list of all available Gateway plasmids can be found here

About qPCR: back to BioRad mix..

Kapa 's SYBR Fast master mix is out..
BioRad 2x iQ SYBR Green Supermix is in again..
Available like before in the bottom drawer in the big Chemicals Room fridge.
The Supermix contains a small amount of fluorescein that permits the collection of well factors on the experimental plate.

In the Chromatin Lab, on the bottom of one of the fridges (the most left-hand one in the ToolBox corner) there is a small blue container with one or two pre-thawed Supermix tubes. Use these tubes when the Supermix in the Chemicals Room fridge is finished; MIX BY INVERTING A FEW TIMES BEFORE USE!!

  May, 25  2011 NEWS:

About qPCR: Kapa mix

Soon we will have ran out of BioRad SYBR Green supermix  for qPCR (in fridge in Chemicals Room, with backup in fridge in Chrom Lab).
Kapa will be our new supplier. Their 2x mix for the BioRad MyiQ qPCR machine is called SYBR Fast mastermix. It will be available in 5 ml tubes, like the BioRad mix.
Read the Product Brochure, Technical Data Sheet, Application Note I and  Application Note II.
The Kapa mix has been well tested by our neighbours of PRI Bioscience.
It is 25% cheaper than BioRad, which still means  about €25 per full qPCR plate…

[you may want to read once more the FD enzymes news of April, 7 2011, below]

 April, 7   2011 NEWS

About the new FD enzymes

At MolBi, we will make the transition to "Fast Digest" restriction enzymes from Fermentas.
The reason is, that Invitrogen appears no longer to be a reliable supplier of REs.

This is the moment to start buying all REs from one-and-the-same supplier, which will be Fermentas.
All enzymes we use are now available from Fermentas, and all are also available as FD enzymes.
In daily use, FD enzymes are 10-20% more expensive than regular enzymes, but have several advantages..:

they cut very fast (may not be very important in most cases..),
but also: all FD enzymes cut in the same buffer (with or without Loading dye; both are supplied..), so double-digestions are no problem!

Enzyme concentration (units per ul) are not given anymore, but all enzymes are titrated so that one microliter digests one microgram DNA in less than 15 minutes.
Since the enzymes are more expensive, a series of test digestions were done at MolBi using only 0.5 ul: perfect result (checked after 30 minutes digestion)..
So, that will be the advice: use 0.5 ul for each digestion. See also the exact reaction conditions info.

After a successful test series using four different FD enzymes samples, transition to FD enzymes has started now, the first three FD enzymes are in stock at MolBi: FD Hpa II, FD Bgl II and FD Rsa I.
They will soon appear in the familiar -20 boxes.
10x FD buffer, and 10x FD Green (includes Loading Dye) can be found in the Fermentas Buffers -20 box.

Unfortunately, this all means, that during the transition, we have five types of enzymes/buffers combinations:

• Invitrogen/REact buffers
• Invitrogen New /H, M, L Buffers
• NEB/NEBuffers
• Fermentas conventional/Fermentas buffers
• FD Fermentas/FD buffers
   

January, 7  2011 NEWS:

Invitrogen supplies new buffers with its Restriction Enzymes (identical to TAKARA enzymes btw..)!!
The REact Buffers are replaced by M, L, H, K and T buffers for recently arrived Invitrogen enzymes
Click here for the new Activity and Buffer Composition chart,
 and here for the new Double Digest Buffer chart.
From now on, the new enzymes and buffers will become available at MolBi.. Enzymes that need the new buffer have an encircled hand-written N indication on the tube label.
The new buffers will be stored in the REact buffers box in the enzymes freezer.

A list of currently (2011_01_07) available enzymes can be found here.
You may need that list to make a MolBi Enzymes Group in ApE.

For qPCR:
BioRad 2x iQ SYBR Green Supermix is available in the bottom drawer in the big Chemicals Room fridge.
The Supermix contains a small amount of fluorescein that permits the collection of well factors on the experimental plate.

In the Chromatin Lab, on the bottom of one of the fridges (the most left-hand one in the ToolBox corner) there are two small aluminum containers with a pre-thawed Supermix tube each. Use these tubes when the Supermix in the Chemicals Room fridge is finished; MIX BY INVERTING A FEW TIMES BEFORE USE!!

Click here for the Supermix Instruction Manual.