Description InstaGene™ Matrix is a Chelex ® -based resin for PCR
Polymerase chain reaction
The polymerase chain reaction (PCR) is a technology in molecular biology used to amplify a single copy or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
How does instagene work?
InstaGene matrix, made with a specially formulated 6% w/v Chelex resin, makes DNA sample preparation fast, easy, and cost-effective, providing PCR-quality template DNA in less than an hour. The Chelex matrix binds to PCR inhibitors rather than DNA, preventing DNA loss due to irreversible DNA binding.
What is instagene matrix used for?
or Instagene Matrix. InstaGene matrix, made with a specially formulated 6% w/v Chelex resin, makes DNA sample preparation fast, easy, and cost-effective, providing PCR-quality template DNA in less than an hour. The Chelex matrix binds to PCR inhibitors rather than DNA, preventing DNA loss due to irreversible DNA binding.
What is the difference between Chelex and instagene?
The Chelex matrix binds to PCR inhibitors rather than DNA, preventing DNA loss due to irreversible DNA binding. The InstaGene matrix eliminates the time-consuming and labor-intensive deproteinization, organic extraction, dialysis, and alcohol precipitation protocols required in traditional DNA purification procedures.
How do I use the instagene microcentrifuge?
Place the cells in a microcentrifuge tube, add the InstaGene matrix, boil, and spin. The matrix adsorbs cell lysis products that interfere with the PCR amplification process.
What is InstaGene and how does it function?
InstaGene matrix, made with a specially formulated 6% w/v Chelex resin, makes DNA sample preparation fast, easy, and cost-effective, providing PCR-quality template DNA in less than an hour. The Chelex matrix binds to PCR inhibitors rather than DNA, preventing DNA loss due to irreversible DNA binding.
What is Chelex resin?
Chelex 100 Resin is a styrene divinylbenzene copolymer containing paired iminodiacetate ions, which act as chelating groups in binding polyvalent metal ions. It can be used for fast and easy DNA and RNA preparations.
How does InstaGene matrix inhibit the activity of dnase?
When DNA is released from your sample in the presense of the InstanGene Matrix, the charged beads grab the Mg2+ and make it unavailable to the enzymes that would degrade the DNA you're trying to extract.
Why is Chelex used in PCR?
Chelex protects the sample from DNases that might remain active after the boiling and could subsequently degrade the DNA, rendering it unsuitable for PCR. After boiling, the Chelex-DNA preparation is stable and can be stored at 4°C for 3–4 months.
What is Chelex made of?
Chelex 100 Resin is made of a styrene divinylbenzene copolymer containing paired iminodiacetate ions, which act as chelating groups in binding polyvalent metal ions.
What is the purpose of the InstaGene Matrix you will add to your DNA quizlet?
What is the purpose of the Instagene matrix? It binds divalent cations, effectively removing them from solution. The cations allow DNA degrading enzymes (DNases) to be active.
What is the reason InstaGene matrix is added to cheek cells?
The cheek cells are transferred to a micro test tube containing InstaGene™ matrix. This particulate matrix is made up of negatively charged, microscopic beads that chelate, or grab, metal ions out of solution. It traps metal ions, such as Mg2+, which are required as catalysts or cofactors in enzymatic reactions.
Why do you think the DNA is stored cold with the InstaGene Matrix?
why do you think dna is stored cold with the instagene matrix after boiling the samples? Keeping the extracted DNA in a refrigerator slow down the activity of any remaining enzymes that could harm DNA and also slow down the bacterial growth.
How do you make a Chelex solution?
10% Chelex Weigh out 1 g of Chelex 100 (100-200 mesh, sodium form from BioRad). Add 50 mM Tris to dry Chelex to make 10 ml of solution. Adjust pH to 11 using 4 N NaOH.
What is Chelex extraction method?
The Chelex method of DNA extraction is suitable for extracting the DNA from a smaller amount of samples. This method is quick and straightforward and does not involve any harmful organic solvents. The basic protocol involves the extraction of DNA by adding samples to hot Chelex suspensions at pH 10–11.
What does Chelex do to the cheek cell extraction?
The Chelex beads will bind divalent magnesium ions (Mg++). These ions often serve as cofactors for nucleases that will degrade your DNA sample and may interfere with the enzyme (Taq polymerase) used in the reaction. By removing magnesium ions, the degradation of genomic DNA by nucleases is reduced.
How do you make a 5% Chelex?
Prepare a 5-10% by weight slurry of Chelex100 Resin (Biorad part 143- 3832,100-200 mesh Chelex, sodium form) and UV sterilized HPLC water. The most effective way to do this is to take a 50 ml sterile falcon tube, place in on a scale inside a small beaker and zero the scale.
Which bacterium has enzymes that are able to withstand the denaturing conditions required during PCR?
Thermus aquaticus bacterium has enzymes that are able to withstand the protein denaturing conditions required during PCR. Because the heating process takes place numerous times in order to make multiple copies, Taq polymerase was isolated form the Thermus aquaticus to be able to withstand the denaturing from the heat.
What are singlue bases?
singlue units of bases A, T, G and C which are essentially building blocks for new DNA strands. They extend an anneal primer. They are called dNTPs or dATP, dTTP, dGTP, dCTP.
What are primers in DNA?
primers are short strands of DNA (usually between 6 and 30 base pairs long) called oligonucleotides. Two primers are needed to amplify a sequence of DNA, a forward and reverse primer. The two primers are designed and synthesized in the laboratory with a specific sequence of nucelotides such that they can anneal (bind) at the opposite ends of th etarget DNA sequence on the complementary strands of the target DNA template. Primers give a specificity to the PCR but they are also necessary because DNA polymerases can only add nucleotides to double stranded DNA. Once the double stranded DNA is separated by heating it, then each primer binds to its complementary sequence on each of the separated DNA strands
