What is a good DNA concentration ng UL?

What is a good DNA concentration ng UL?

The effective read range of UV spectroscopy is 0.1 to 0.999 which corresponds to approximately 4 ng/µL to 50 ng/µL of genomic DNA. Values above or below that range are invalid absorbance readings.

How do you quantify ssDNA?

The ssDNA Quantification Kit is developed for single stranded DNA quantification. The kit includes ssDNA Dye, ssDNA Dilution Buffer, and two ssDNA Standards. Simply dilute the ssDNA Dye with the ssDNA Dilution Buffer, add DNA sample (volume from 1-20 μL), then read the concentration using the Qubit® Fluorometer.

How do you calculate dsDNA and ssDNA concentration from the A260 value?

A260 as quantity measurement One A260 unit is the amount of nucleic acid contained in 1 mL and producing an OD of 1. The same conversion factors apply, and therefore, in such contexts: 1 A260 unit dsDNA = 50 µg. 1 A260 unit ssDNA = 33 µg.

What is a high DNA concentration?

for DNA sizes above 500bp, it is recommended the minimum concentration is 40ng/ul with a minimum volume of 15uL. for sizes below 500bp, 20ng/uL is sufficient. I have tried sending in samples as low as 10ng/uL for above 500bp range and still get back some sequencing data.

What is a normal DNA concentration?

A ratio of ~1.8 is generally accepted as “pure” for DNA; a ratio of ~2.0 is generally accepted as “pure” for RNA. If the ratio is appreciably lower in either case, it may indicate the presence of protein, phenol or other contaminants that absorb strongly at or near 280nm.

What is an acceptable 260 280 ratio for DNA?

The ratio of absorbance at 260 and 280 nm is used to assess DNA purity. A ratio of ∼1.8 is generally accepted as “pure” for DNA. If the ratio is appreciably lower (≤1.6), it may indicate the presence of proteins, phenol, or other contaminants that absorb strongly at or near 280 nm.

What is the meaning of ssDNA?

single-stranded DNA
ssDNA (plural ssDNAs) single-stranded DNA. (virology) A group of viruses with such DNA.

How accurate is NanoDrop?

At higher dilutions (sixteen fold and above, DNA concentration range: 1.35–0.16 ng/μl), quantification by NanoDrop was neither precise nor accurate, while quantification by Qubit was found to be precise and close to estimated concentration of DNA at the tested dilution.

What does the 260 230 ratio mean?

260/230 Ratio This ratio is used as a secondary measure of nucleic acid purity. The 260/230 values for “pure” nucleic acid are often higher than the respective 260/280 values. Expected 260/230 values are commonly in the range of 2.0-2.2.

What is a good DNA concentration for PCR?

25 -100 ng
Genrally 25 -100 ng human genomic DNA is recomended for PCR. So around 10,000 – 12000 copies of target DNA are recomended in 25 ul pCR reaction……..

How do you know if DNA is contaminated?

To check solutions for contamination, assemble negative control reactions using new reagents known not to be contaminated, and add one of the suspect solutions to each reaction. A reaction that shows amplified products indicative of contamination is evidence that the solution added was contaminated.

Why does ssDNA have a higher 260 280 ratio than dsDNA?

ssDNA degrades faster, thus absorbance increases at 260nm ssDNA folds to become double stranded, which increases absorbance sDNA is single-stranded, thus the nitrogenous bases are presented SSDNA is single-stranded, thus the sugar-phosphate backbone can absorb better at 260nm.

Why is my 260 280 ratio so high?

Abnormal 260/280 ratios usually indicate that the sample is either contaminated by protein or a reagent such as phenol or that there was an issue with the measurement. High 260/280 purity ratios are not indicative of an issue.

What does A positive ssDNA mean?

A high level of anti-dsDNA in the blood is strongly associated with lupus and is often significantly increased during or just prior to a flare-up. When the anti-dsDNA is positive and the person tested has other clinical signs and symptoms associated with lupus, it means that the person tested likely has lupus.

What is ssDNA test?

Anti-ssDNA is an ELISA-based, automated, in-vitro test system for the quantitative, in-vitro determination of IgG antibodies against single-stranded DNA in human serum or plasma. This assay is used for the serological detection of systemic lupus erythematosus (SLE) and drug-induced lupus.