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Question 1

-Pass criteria (concen. 50ng)
Array-20ng
Dpyd-10ng
260/280 is between 1.75 and 2.04
260/230 is 1.5 and 3.5 absorbance

280 is protein contamination
260 is DNA
230 is reagents I.e. Phenol

Qubit
Picogreen
Nanodrop (wavelengths)

Sample below 50 needles re-extract

Ethanol precipitation

Clean up (scientist decision)

How to calculate concentration.

Question 2

Picogreen

Answer 2

is a specialized tool used in molecular biology for accurately measuring the quantity of double-stranded DNA (dsDNA) in a sample.

Question 3

Here’s a breakdown of what Picogreen is and how it’s used:

Answer 3

Function:
Picogreen is a highly sensitive fluorescent nucleic acid stain. This means it binds specifically to double-stranded DNA and emits light (fluoresces) when exposed to a specific wavelength.
The amount of fluorescence emitted is directly proportional to the amount of dsDNA present in the sample. By measuring the fluorescence intensity, scientists can determine the concentration of dsDNA.

Question 4

Advantages of Picogreen:

Answer 4

Sensitivity: Picogreen is significantly more sensitive than traditional methods like absorbance at 260 nm (A260), which can be influenced by contaminants. It allows researchers to measure very small amounts of DNA, as low as 50 picograms (pg) per milliliter (mL).

Specificity: Picogreen binds specifically to dsDNA, with minimal interference from single-stranded DNA (ssDNA), RNA, or other common contaminants in biological samples. This provides a more accurate measurement of double-stranded DNA concentration.

Ease of Use: The Picogreen assay is relatively simple and can be performed in a microplate reader or fluorometer.