Bst Large Fragment

Bst Large Fragment

Reagent Sharers

Alex Brown


This enzyme comes in a pET15b plasmid. Information shared by Alex: This plasmid contains a DNA Polymerase which retains 5´ → 3´ polymerase activity from full length Bst DNA Polymerase, while lacking 5´ →3´ exonuclease activity. The polymerase has been described for use in applications requiring thermophilic strand displacement. Used for: Loop-Mediated Isothermal Amplification (LAMP), DNA sequencing through high GC regions, rapid sequencing from nanogram amounts of DNA template.

This enzyme is a codon-optimized version of Andy Ellington´s Bst LF

Expression/Production Information (if applicable)

Protocol Information

Expression and purification protocol currently available at ReClone
group in

How to get the plasmid/material/reagent

Currently in Alex´s hands, FreeGenes and Chile/Argentina nodes
This is vector received from Alex and being used in Chile.


Under openMTA from Alex, being able to redistribute under openMTA from iBio/Chile too.


(PMID: 10871386, 25652028, 1793578)

Hi everyone! Is this polymerase available for commercial use?

Yes! It is distributed under the OpenMTA which explicitly allows commercial use. The Bst-LF US patent expired in 2016 so it is in the public domain.

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Javier, do you have a protocol to share in for BstLF?
(all protocols here:

I hope to have one by the end of the week. Based on Alex Brown’s protocol

Hi, please be aware the original post :point_up_2: has been edited to include the expression and purification protocol (

We are currently working on a protocol for RT-LAMP based on this enzyme, MMLV and in-house prepared buffers. Preliminary results indicate that 10.7 ng/uL of BstLF (this is uL of reaction so 267.5 ng of BstLF total in a 25 uL reaction volume) is equivalent to the performance of 8U of commercial Bst2.0 (and around 125 pg/uL of MMLV, again uL of reaction so 3.1 ng of MMLV total in a 25 uL of reaction volume). We use the N2 primer set for the characterization and T7-based in vitro transcribed gene N (DNA from IDT) as a target. We used in-house prepared reaction buffers based on NEB 1X Isothermal Amplification Buffer Pack. Isaac Nuñez and Tamara Matute are now working on the optimization of these concentrations. Collabs/ideas are very welcome.


Hi! My lab in the north of Brazil recently engaged to explore the RT-LAMP technique using commercially available enzymes and now we want to establish in house production. Would you please indicate where can I find this vector (is it available in AddGene)? Thanks

Hi Jimmy, yes we can send you some DNA.
Could you please send us your address?

That would be really kind! May we get your email to send the informations?


Hi Jimmy,
Ariel here, I work with Fernan and I can help you with some DNA

Write us at ariel.cerda.rojas(at)gmail(dot)com.


Hi All, I recently saw this new preprint where they used drying (without freezing) for an RT-LAMP reaction, and thought it might be interesting for some of you,

Not sure if this is the right place or maybe we should have a channel for Lyophilizatoin/Stability/Storage/Shipping Issues?
Happy Holidays!

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