Check Know the purposes of using a micro spin column and SYBR Green qPCR Mix

 

A micro spin column plays a vital role in purifying deoxyribonucleic acid quickly. The purified DNA can be used for desalting it, exchanging the buffer, as well as for eliminating un-integrated nucleotides from end-labeled oligonucleotides. These spin columns are the tools to manipulate small volumes of affinity supports conveniently, usually between 5 µLs and 100 µLs for purifying proteins.

The major benefit of using a micro spin column during the process of affinity purification is that it will purify more proteins in less time. These spin columns are intended to purify DNA rapidly when they are used together with Sephadex G-50 DNA Grade or G-25 DNA Grade. Both forms of Sephadex are highly suitable to purify oligonucleotides or very small volumes of deoxyribonucleic acid following mixture or a labeling reaction.

The major reason for using these two grades of Sephadex is that Sephadex G-50 is a deep-rooted gel filtration resin. It is used for desalting as well as for buffer swap of biomolecules with a molecular weight of more than 30 000. On the other hand, Sephadex G-25 is one of the five diverse G-types, varying from G-10 to G-75. While G-10 is mostly used for excluding small molecules, G-75 is used for larger molecules. Moreover, it has an elimination limit of about Mr 5000.

Micro spin columns are also quite useful in desalting or exchanging PCR products as well as other DNA specimen in a volume of 10 µL to 100 µL by making use of spin-column chromatography. They are the outstanding tools to purify freshly synthesized oligonucleotides of more than 10-mers in a deprotection solution with a volume that ranges from 100 µLs to 150 µLs. They are extremely flexible to use for experiments, meaning they will allow varying amounts of Sephadex. These spin columns can be used for other purposes, as well. Both grades of Sephadex are sold separately.

When it comes to the applications an SYBR Green qPCR Mix, it is mostly used for a real-time Polymerase chain reaction. This is for the reason that it is considered a beneficial, flexible, and easy-to-use gene expression master mix. Moreover, the mix consists of antibody-arbitrated Taq DNA polymerase with a hot-start device, offering tight control over the Taq enzyme start and assisting in preventing unwanted early polymerase activity at low temperatures. Some of the other beneficial features of the mix include:

·         The bi-color tracking dye system of the mix shows the point of pipetting.

·         The mix is compatible with broad primer concentration and primer Tm, allowing greater flexibility while setting up for qPCR reaction with minimum optimization.

·         The mix has a better specificity as well as taut reproducibility in the values of Ct over a wide energetic array to improve data quality.

·         The mix is capable of giving quick and reproducible results, as it works well with the SuperScript IV VILO master mix.

·         The mix has been incorporated with dUTP/ UNG to prevent infectivity of the reschedule PCR products.

Above all, the SYBR Green qPCR Mix is renowned for its high instrument compatibility. Moreover, the tracking dye of the mix aids greatly in reducing pipetting errors.