MCBLab \\ Spectrometry

What is Spectrometry?

describes the amount of light absorbed by particles & measure the particles in a concentration

The colors we see are refracted from light?

true

What are DNA and Proteins absorbed at?

DNA- 260 nm Proteins- 280 nm

What is the importance of the LAMD beer law?

tells you how much concentration is being abosrbed

What is the formula for LAMD beer Law?

A= E D C

what is “e” (m) in lamd beer law

the extinction cofactor (L,mol); slope

what is the “d” (m) in lamd beer law

distance the light travels (cm)

what is “c” (x) in lamd beer law?

cocentration of abosrbing solution(mol/L)

What is Fast green?

a dye used to bind protein absorbed at 625 nm

highest peak= highest aborbance T/F?

true

In a grpagh, what is the indepednat varible and dependant for OD vs concentration?

OD @ 625= dependant; Concentration (uM)= independant

What is the DNA purity formula?

OD260/OD280

if 1.8 or more? DNA

PURE DNA sample, no protein present

if 1.8 or less? DNA

Contaiminated protein in DNA sample

purpose of serial diluations?

if the OD valye is above 1.0 , you pick a small portion of stiock, and diilute the rest with DI water for OD value to drop- multiply OD value by dilution factor

formula for number of mols?

#mols= M*mm

Dilution Formula

C1V1= C2V2

what happens if you dilute soemthing?

it reduces the amount of solute in a soltuion

DNA Cocentration Formula

50ug/mL *OD

What is the measuremnt to go from Liters—> mL, and mL—→liters

Liters—> mL *1000, mL—→liters /1,000

What is the measuremnt to go from uL—> mL, and mL—→uL

uL—> mL /1,000, mL—→uL *1,000

What are the ranges for pippetes P2, P20, P200, P,1000

• P2: 0.2 – 2 µL

• P20: 2 – 20 µL

• P200: 20 – 200 µL

• P1000: 100 – 1000 µL (1 mL)

OD has units? T/F

false

cocnentration formula basic

C=mols/ volume

10% solute

10 grams of solute +100 mL of DI water

20% cocentraion

20 grams of cocentration + 80 mL of DI water

if higher light is omitted less is transmitted? t/f

True

acid/ base features?

• acid or base

• Ph= 7 →neutral 

• Ph = 1-6→ acidic (lowers ph)

• ph = 8-14→ basic 

whats tris?

the buffering agent

spectrometer

determines the amount of light of a particle “color” or wavelneght that is ABSORBED by particles