Bst DNA Polymerase (large fragment)

¥0.00¥35,094.68

For DNA sequencing high GC regions and rapid sequencing from nanogram amounts of DNA templates.
Bst.jpg"
SKU: BPL 分类: ,
SKU OPTIONS 价格
BPL-100 Bst DNA Polymerase (large fragment), 8,000 units (8 U/µl) ¥1,451.00
BPL-200 Bst DNA Polymerase (large fragment), 10,000 units (100 U/µl) ¥1,547.00
BPL-300 Bst DNA Polymerase (large fragment), 50,000 units (100 U/µl) ¥3,153.00
BPL-400 Bst DNA Polymerase (large fragment), 100,000 units (100 U/µl) ¥6,305.00
BPL-500 Bst DNA Polymerase (large fragment), 1,000,000 units (100 U/µl) ¥35,095.00
BPL-800 Bst DNA Polymerase (large fragment Glycerol-Free), 10,000 units (400 U/µl) ¥1,675.00
BPL-900 Bst DNA Polymerase (large fragment Glycerol-Free), 100,000 units (400 U/µl) ¥6,814.00
BPL-1000 Bst DNA Polymerase (large fragment Glycerol-Free), 400,000 units (400 U/µl) ¥26,565.00
B-PB10 10x PCR Buffer II (1ml) ¥173.00
BPL-OEM Bst DNA Polymerase (large fragment) (Any Size) Please inquire

描述

Description:
Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the 5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.

Enzyme provide strong strand displacement activity. The optimum temperature is from 60-65 ⁰C. Bst becomes heat inactivated at 80 ⁰C. Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a gene of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain.

Bst DNA Polymerase, large fragments is available in three concentrations, 8,000 U/ml, 50,000 U/ml and 400U/ml in a storage buffer of 10 mM Tris-HCl, pH 7.5, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X-100, and 50% Glycerol. High concnetration (400U/ml) also is supplied in glycerol free buffer and lyophilized powder.
Bst.jpg"
Figure: MCLAB Bst DNA Polymerase (large fragment) shows higher strand -displacing polymerase activity than two other vendors’ products.

 

Application:

– nucleic acid amplification methods, including isothermal amplification, LAMP
– whole genome amplification
-Sequencing through problematic secondary structures

Typical LAMP Protocol with Bst DNA Polymerase

Component Final Concentration 25 µl Reaction
10X Buffer for Bst 1X (does not contain MgSO4) 2.5 µl
MgSO4 (100 mM) 8 mM total 2,0 µl
dNTP Mix (10 mM) 1.4 mM each 3.5 µl
FIP/BIP Primers (10X) 1.6 µM 1 µl
F3/B3 Primers (10X) 0.2 µM 1 µl
LoopF/B Primers (10X) 0.8 µM 1 µl
Bst Large fragment (8,000 U/ml) 80-320 U/ml 0.95-1.1 µl
DNA Sample > 10 copies or more NA
Nuclease-free Water up to 25 µl
Total Reaction Volume 25 µl

 

Specific Activity: 120,000 U/mg

Supplied in:
10 mM Tris-HCl
50 mM KCl
1.0 mM DTT
0.1 mM EDTA
0.1% Triton X-100
50% Glycerol
pH 7.5 @ 25°C
Supplied with:
10X PCR buffer II

10x PCR Buffer ll:
200 mM Tris-HCl
100 mM Ammonium Sulfate
100 mM KCl
20 mM MgSO4
1.0% Triton X-100
pH 8.8 @ 25°C

Unit Definition:
1 unit is defined as the amount of polymerase required to convert 10 nmol of dNTPs into acid insoluble material in 30 minutes at 65°C.

Recommended Storage Condition: -20°C

Note:

– Recommended for long term storage: 0.1% Triton X-100
– Reaction temperatures above 70°C are not recommended.
– Bst DNA Polymerase cannot be used for thermal cycle sequencing.

Reference:
Kiefer, et al. Structure 15 January 1997. 5, 95-108.

其他信息

OPTIONS

Bst DNA Polymerase (large fragment), 8,000 units (8 U/µl), Bst DNA Polymerase (large fragment), 10,000 units (100 U/µl), Bst DNA Polymerase (large fragment), 50,000 units (100 U/µl), Bst DNA Polymerase (large fragment), 100,000 units (100 U/µl), Bst DNA Polymerase (large fragment), 1,000,000 units (100 U/µl), Bst DNA Polymerase (large fragment Glycerol-Free), 10,000 units (400 U/µl), Bst DNA Polymerase (large fragment Glycerol-Free), 100,000 units (400 U/µl), Bst DNA Polymerase (large fragment Glycerol-Free), 400,000 units (400 U/µl), 10x PCR Buffer II (1ml), Bst DNA Polymerase (large fragment) (Any Size)

1. Does Bst DNA Polymerase have a 3′->5′ proofreading exonuclease activity?
No.

2.What is the difference between Bst DNA Polymerase (regular) and Bst DNA Polymerase, large fragment?
Bst DNA Polymerase, large fragment does not have the N-terminal exonuclease domain, so it not only lacks 3′->5′ exonuclease activity, it also lacks 5′->3′ exonuclease activity.

3. What is the best temperature range for Bst DNA Polymerase to be used?
The best temperature range is 60-65°C, it has 100% activity. The enzyme also has activity

4.  What are the benefits of choosing Bst DNA Polymerase?
Bst DNA Polymerase, either regular or large fragment, is good at strand displacement with optimal temperature of 60-65°. This gives researchers a wider range of reaction conditions to optimize strand displacement and primer annealing. This is useful in the design of sequencing strategies as well as isothermal amplification technologies. The elevated reaction temperature facilitates sequencing through GC rich regions.

5. Can Bst DNA Polymerase be used for thermal cycle sequencing?
No, the reaction temperatures are too high for enzyme stability. It becomes inactive at 80°C for 10 minutes.