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A Treasure Box of Tools for Genetic Engineering

6) Cloning and Gene Analysis


Products on this page (Cloning and Gene Analysis)

  • MagExtractor -PCR & Gel Clean Up | high efficient DNA fragment purification kit
  • Ligation high Ver.2 | Ready-to-use ligation reagent
  • TArget Clone | high efficient TA cloning kit
  • TArget Clone -Plus-  | high efficient TA cloning kit for KOD products
  • 10xA-Attachment Mix | dA attachment reagent
  • KOD -Plus- Mutagenesis Kit | site-directed mutagenesis kit
  • PerfectHyb |high efficient hybridization solution
  • 100 bp DNA Ladder
  • 1 kb DNA Ladder
Cloning and Gene Analysis Applications Product Name
DNA fragment purification MagExtractor -PCR & Gel Clean up-
High efficient ligation Ligation high Ver.2
TA cloning of PCR products having overhanged dA at the 3'-ends. TArget Clone
PCR products from Taq DNA polymerase, Blend Taq [Cat. No. TYB-BTQ-101],
Blend Taq -Plus- [Cat. No. TYB-BTQ-201] or KOD Dash [Cat. No. TYB-LDP-101]
are available.
TA cloning of blunt-end PCR products amplified using TArget Clone -Plus-
KOD -Plus- [Cat. No. TYB-KOD-201] or KOD FX [Cat. No. TYB-KFX-101].
Attachment of dA at the 3' - ends of blunt end PCR products 10xA-attachment mix
from KOD -Plus- and KOD FX ( For TA cloning )
Site-directed mutagenesis KOD -Plus- Mutagenesis Kit
Southern & Northern hybridization PerfectHyb hybridization solution
100 bp DNA Ladder Marker 100 bp DNA Ladder
1 kb DNA Ladder Marker 1 kb DNA Ladder

tyb-small-logo-inline.pngCloning & Gene Analysis Products

DNA FRAGMENT PURIFICATION 

high efficient DNA fragment purification kit
MagExtractor -PCR & Gel Clean up-  

  • Extracts DNA fragments from ≤ 100 μl PCR solutions or enzyme solutions (e.g., restriction enzyme and alkaline phosphatase) within 5 minutes.
  • Extracts DNA fragment from ≤ 0.3 g agarose gel slices (TAE or TBE) within 15 minutes. Agarose slices can be melted at room temperature; it is not necessary to use a heating block.
  • Typical yields from solution or gel slices are approx. 60-70%. DNA fragments approx. 100 bp to 50 kb can be effectively recovered. Small fragments (< 40 bp) are removed.
  • Purified DNA fragments can be applied to sequencing, restriction enzyme treatment, labeling, ligation, transformation, etc. RI-labeled probes can also purified with this kit.

MagExtractor -PCR & Gel Clean up- provides a simple and reliable method for the rapid purification of DNA fragments from a PCR solution, enzyme solution, or agarose gel slices utilizing magnetic silica beads. This kit is based on binding properties of DNA to a silica surface in the presence of chaotropic agents. The purified DNA fragments can be used directly for general molecular biology experiments.

 HIGH EFFICIENT LIGATION 

ready-to-use ligation reagent
Ligation high Ver.2  

  • Effective ligation of cohesive, blunt and A overhang DNA fragments can be achieved.
  • Will not freeze at -20ºC. No need to thaw.
  • Just mix ligation high Ver.2 with an equal volume or with double the volume of the solution containing DNA fragments.

The ligation reaction is an essential step in genetic recombination experiments. For this reaction, T4 DNA ligase has been widely used. Ligation high Ver.2 is a highly efficient premixed T4 Ligase reagent.

Reagents at -20ºC
Left: conventional, Right: Ligation high Ver.2

HIGH EFFICIENT CLONING 

high efficient TA cloning kit
TArget Clone 

  • Ligation step can be completed in 5 min with high efficiency.
  • PCR products can be used without purification.

TArget Clone [Cat. No. TYB-TAK-101] can be applied to the TA cloning of PCR products amplified using Taq DNA polymerase, Blend Taq [Cat. No. TYB-BTQ-101], Blend Taq -Plus- [Cat. No. TYB-BTQ-201] or KOD Dash [Cat. No. TYB-LDP-101]. Almost all PCR products having dA overhanging at the 3'end are available.

high efficient TA cloning kit for KOD products
TArget Clone -Plus-  

  • Can be applied to the TA cloning of blunt-end PCR products of KOD DNA polymerase.
  • Ligation step can be completed in 5 min with high efficiency.
  • PCR products can be used without purification.

TArget Clone -Plus- [Cat. No. TYB-TAK-201] can be applied to the TA cloning of blunt-end PCR products amplified using KOD -Plus- [Cat. No. TYB-KOD-201] or KOD FX [Cat. No. TYB-KFX-101]. TArget Clone -Plus- contains 10xA-attachment mix. This reagent is a mixture of anti-KOD DNA polymerase antibody specific to KOD 3'→5' exonuclease activity (proof-reading activity) and Taq DNA polymerase, which exhibits terminal transferase activity. The 10 x A-attachment mix allows for blunt end PCR products to acquire overhanging dA at the 3'-ends

TA CLONING OF BLUNT-END PCR PRODUCTS OF KOD SERIES 

dA attachment reagent
10xA-attachment mix 

  • The attachment reaction is completed in 10 minutes at 60ºC.
  • The cloning efficiency of dA attached PCR products is as great as that of PCR products of Taq DNA polymerase.

10 x A-attachment mix is a reagent consists of the anti-KOD DNA polymerase antibody for the 3'→5' exonuclease activity (proof-reading activity) of the enzyme, and Taq DNA polymerase exhibits the terminal transferase activity. The PCR products from KOD -Plus- [Cat. No. TYB-KOD-201] and KOD FX [Cat. No. TYB-KFX-101] have blunt ends due to the 3'→5' exonuclease activity of KOD DNA polymerase. 10 x A-attachment mix allows the PCR products to acquire overhanging dA at the 3'-ends. The products having 3'-dA overhang can be directly cloned into arbitrary T-vectors using ligation reagents such as Ligation high Ver.2 [Cat. No. TYB-LGK-201].

SITE-DIRECTED MUTAGENESIS 

site-directed mutagenesis kit
KOD -Plus- Mutagenesis Kit  

  • Applicable for various mutations, such as substitutions, insertions, and deletion mutations.
  • High efficiency (95% maximum) can be obtained.
  • Simple protocol facilitates speedy experiments. Phosphorylated primers are not required.

>KOD -Plus- Mutagenesis Kit is an inverse PCR (iPCR)-based site-directed mutagenesis kit using KOD DNA polymerase as a high-fidelity PCR enzyme. This reagent was developed based on a highly efficient PCR reagent, "KOD-Plus- (Cat. No. TYB-KOD-201)", which consists of KOD DNA polymerase and anti-KOD DNA polymerase antibodies for Hot Start PCR. This kit enables not only the introduction of point mutations, but also the introduction of large insertions and deletions. The PCR fidelity of KOD-Plus- is greater than Taq DNA polymerase (ca. 80-fold); therefore, unexpected, 2nd-site mutations can be reduced. PCR reactions can be performed using standard PCR primers and do not require phosphorylated primers, because this protocol contains a 'Phosphorylation Step' of PCR products.

SOUTHERN & NORTHERN HYBRIDIZATION 

high efficient hybridization solution
PerfectHyb 

  • Efficient and sensitive RI and non-RI detection by Southern and Northern blotting analyses.
  • Reduces hybridization time from 16 to 1-2 hr.
  • Over night hybridization significantly increases the sensitivity on detection of low levels of expression mRNA by Northern blotting analysis.
  • Low viscosity and no generation of precipitates at room temperature.
  • Addition of salmon sperm DNA is necessary.
  • Hybridization and washing steps can be performed at the same temperature.

PerfectHyb hybridization solution contains an accelerator that improves the hybridization efficiency and background. The hybridization rate is accelerated and background hybridization minimized. The solution facilitates Southern and Northern blotting analysis.

DNA LADDER MARKERS

100 bp DNA Ladder 

  • Dye-attached type and dye-mixed type (Loading Quick™) are available.
  • The 500 bp fragment is enhanced.
  • The DNA mass in each band is determined.

A 100 bp DNA Ladder is suitable for use as a molecular weight standard (100 bp-1,500 bp) for agarose gel electrophoresis. A 500 bp fragment has increased intensity as a reference band. Dye-attached type and dye-mixed type (Loading Quick™) are available.

1 kb DNA Ladder 

  • Dye for electrophoresis is attached to the product.
  • The 5 kb fragment is enhanced.

A 1kb DNA Ladder is suitable for use as a molecular weight standard (1,000 bp-10,000 bp) for agarose gel electrophoresis. A 5,000 bp fragment has increased intensity as a reference band.

Product name Code No. Packaging storage temperature Manual specifications SDS
dNTPs Set NTP-101 0.5mL x 4 -20 ℃ D ―
dNTPs Mixture NTP-201 1.0mL x 1 -20 ℃ D S
(A, C, G, T each 2mM)
dNTPs Mixture NTP-501 1.0mL x 1 -20 ℃ M D S
(A, C, G, U each 2 mM)
dNTPs Mixture NTP-301 0.2mL x 1 -20 ℃ D S
(A, C, G, T each 10mM)
dUTP (100 mM) UTP-101 0.5mL x 1 -20 ℃ M D S