O slideshow foi denunciado.
Utilizamos seu perfil e dados de atividades no LinkedIn para personalizar e exibir anúncios mais relevantes. Altere suas preferências de anúncios quando desejar.
Calculating the size of DNA fragments
using unknown marker length
Nishanth Sekar
PhD (Ag) in Microbiolgy
Problem:
Calculate the size of A, B, C using the Hind III digest of lamba DNA marker
Create standard curve using DNA marke...
Create standard curve using DNA marker
Size of DNA fragment in
bp
log 10
Distance travelled by
the fragments (cms)
23130 4...
Measure distance travelled by unknown
DNA fragment
Distance travelled by A = 6.5 cm
Distance travelled by B = 10.7 cm
Dist...
Determine size of DNA fragment
A
B
6.5 cm 10.7 cm 13.7 cm
2.7
3.2
3.8
A : antilog (3.8) = ~ 6309 bp
B : antilog (3.2) = ~ ...
Using MS Excel
Simple linear regression equation
y = mx+c
In this case…
y = -0.149x+4.792
Size of DNA fragment in
bp
log 1...
How to calculate the size of DNA fragments using unknown marker length
Próximos SlideShares
Carregando em…5
×

How to calculate the size of DNA fragments using unknown marker length

The size of DNA fragment or the gene of interest could be estimated using standard markers (like 1kb, 100bp markers etc.). If our fragment run between the known marker fragment, then this method is useful.

Livros relacionados

Gratuito durante 30 dias do Scribd

Ver tudo
  • Seja o primeiro a comentar

  • Seja a primeira pessoa a gostar disto

How to calculate the size of DNA fragments using unknown marker length

  1. 1. Calculating the size of DNA fragments using unknown marker length Nishanth Sekar PhD (Ag) in Microbiolgy
  2. 2. Problem: Calculate the size of A, B, C using the Hind III digest of lamba DNA marker Create standard curve using DNA marker Measure distance travelled by unknown DNA fragment Determine size of DNA fragment Size of unknown A, B, C ? !
  3. 3. Create standard curve using DNA marker Size of DNA fragment in bp log 10 Distance travelled by the fragments (cms) 23130 4.3642 4.4 9416 3.9739 5.0 6557 3.8167 5.9 4361 3.6396 7.1 2322 3.3659 9.2 2027 3.3069 9.8 564 2.7513 14.8 125 2.0969 17.5
  4. 4. Measure distance travelled by unknown DNA fragment Distance travelled by A = 6.5 cm Distance travelled by B = 10.7 cm Distance travelled by C = 13.7 cm
  5. 5. Determine size of DNA fragment A B 6.5 cm 10.7 cm 13.7 cm 2.7 3.2 3.8 A : antilog (3.8) = ~ 6309 bp B : antilog (3.2) = ~ 1585 bp C : antilog (2.7) = ~ 501 bp C
  6. 6. Using MS Excel Simple linear regression equation y = mx+c In this case… y = -0.149x+4.792 Size of DNA fragment in bp log 10 Distance travelled by the fragments (cms) 23130 4.3642 4.4 9416 3.9739 5.0 6557 3.8167 5.9 4361 3.6396 7.1 2322 3.3659 9.2 2027 3.3069 9.8 564 2.7513 14.8 125 2.0969 17.5 Distance travelled (cms) Value of y Antilog A 6.5 3.8235 6660.395 B 10.7 3.1977 1576.522 C 13.7 2.7507 563.2483 Trendline !!! Size of A, B and C

×