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Genotyping kit, protocol

Protocol and method for quickly generate clean genotyping PCR from un-purified DNA of ear punch, tail, or ear of mice and rats

Neuvitro Corporation owns the leading technology to provide professional coating with the broadest coverage of substrates, such as PDL, PLL, PLO, Collagen, Gelatin, Laminin, Fibronectin, PDL+Laminin, PDL + Fibronectin, PLO+Laminin, PLO+Fibronectin, PDL+Laminin+Fibronectin, etc. The majority of coated German coverslips available from Electron Microscopy Science, ThermoFisher Scientific, and VWR is actually manufactured by Neuvitro Corporation.

GDE-250, the simplest genotyping reagent of genomic DNA extraction kit:

Neuvitro genotyping kit provides the easiest way to extract DNA from mouse and rat tissues, such as a tiny small piece of toes, tails, or ear punch. No purification is required for high quality PCR results.

When open the genotyping kit

Before using the kit, the reagents should be diluted as follows:

  1. Transfer the Reagent A into a 15ml tube and add sterile dd-water to final volume of 15ml.

  2. Transfer the Reagent B into another 15-ml tube and add sterile dd-water to final volume of 15ml.

  3. Store diluted 1x Reagents A and B at room temperature for daily usage. Expiration date: 2 month after dilution. Be careful to mark the two tubes clearly as A or B to avoid mistakes. Undiluted stock can be stored at dark Room Temperature for half year.

Operation procedure:

  1. Collect tiny small pieces (about 1- 1.5mm³) of mouse tissues into 0.5ml micro-tubes (or particular micro-tubes fitting to your PCR machine holes).

  2. Add A into each tube.  Make sure the caps are securely sealed and the tissue is immersed in Reagent A instead of sticking on tube wall (leaking tube during heating will result in failure).

  3. Place tubes into PCR thermocycler and run a program of 94°C for 1 hour.

  4. Add B to each tissue tube right after the 1 hour extraction.  Do not store the samples without addition of reagent B.

Notes:

The tissue remains visible after 1 hour heated DNA extraction. Do not vortex the samples. Take DNA samples from clear supernatant for Genotyping PCR. For 20ul PCR reaction, 1ul of the DNA sample should generate strong bands. But you can adjust sample volume 0.5ul – 2ul if desired.

DNA Storage: 

Keep the DNA samples 4°C short-term before genotyping PCR, then -20°C long-term after PCR.

Collection of genomic DNA extraction kit:

Promega

Maxwell 16 mouse tail genotyping DNA purification kit

G Biosciences

Omniprep genotyping DNA extraction kit from mouse tail

Analytik Jena

Blackprep Genotyping DNA kit for odent tail

Qiagen

Gentra Puregene mouse tail kit for isolate genomic DNA

Xpressbio

nexttec genomic DNA isolation kit from mouse tail

Collection of genotyping kit

EZ bioresearch

New Extraction Protocol:

1) Add DNA Extraction Buffer to the Strip tube with a tail sample.

2) Place the strip tube in a thermocyler and start the extraction program

3) After 20-min, take out the strip tube and invert 5 times.

 

Extraction is done and genomic DNA is ready for PCR.

Features and Benefits

Automated extraction:

Fast Protocol:

Multiplexing:

Easy Handling:

Significant savings:

Consistent results:

Push-button DNA extraction with <10-second hands-on time.

Genomic DNA ready for PCR in 20 min. No overnight digestion

Strip tube for parallel processing

One solution extraction. No termination buffer

No need to purify genomic DNA with expensive spin columns.

Virtually 100% success rate!

Quanta bio

Features & Benefits

  • Simplified, completely reagent-based system requires minimal pipetting skill

  • Premixed electrophretic mobility loading dye reduces chances for post-PCR cross contamination

  • Stabilized 2X PCR SuperMix enables convenient room-temperature setup and is unaffected by repetitive freeze-thaw (>20X)

  • High-yielding, ultrapure modified Taq DNA polymerase delivers robust, reliable duplex assay performance

  • Stringent, ultrapure antibody hotstart ensures sensitive and specific target amplification

 

AccuStart II PCR Genotyping Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

The AccuStart II Genotyping Kit is a complete reagent kit designed to support conventional, end-point PCR-based screening of transgenic animal models commonly used in life science research and is validated for use with mouse, fish, or insect tissue specimens. It combines a rapid, 2-component DNA extraction reagent with a user-friendly 2X concentrated PCR SuperMix with loading dye for seamless gel electrophoresis analysis. qPCR-grade genomic DNA template is obtained with minimal extraction volumes (≤ 100uL) and can be carried out in ≤ 30-minutes on a standard PCR thermal cycler.

Azura

Azura™ Mouse Genotyping Kit provides a rapid, low-cost and highly efficient method for the DNA extraction and amplification of a variety of solid tissue types, and is ideally suited to mouse tail snips and mouse ear punches. Conventional methods require laborious, time-consuming and inefficient steps such as phenol extractions and overnight incubations.

Azura™ Mouse Genotyping Kit includes a proprietary lysis buffer formulation which delivers a rapid and complete release of genomic DNA in 15 minutes, followed by PCR amplification using Azura 2x HS Red Mix for unrivalled sensitivity and performance regardless of sequence complexity. Azura 2x HS Red Mix also includes an inert red dye which facilitates direct gel-loading.

 

Creative Biogene

Creative Biogene Mouse Tail Direct PCR Kit is specially designed for amplification of DNA from mouse tail. This kit contains a novel buffer system that effectively lyses tissue and neutralizes inhibitors. No DNA purification is required and unpurified DNA can be directly used as template for PCR.

Applied Stem Cell

The TARGATT™ Mouse Genotyping Kit allows for convenient genotyping of TARGATT™ Transgenic Mice via tail biopsy. The kit contains reagents, PCR primer sets, buffer along with detailed protocols and anticipated results. This kit is intended for use after TARGATT™ Transgenic mouse model generation using the TARGATT™ Transgenic Kit, TARGATT™ Plasmids, and TARGATT™ “attP” Mice to confirm the transgene integration into the TARGATT™ attP docking site at either the mHipp11 (mH11) or mRosa26 locus.