pcr: PCR Setup Calculator

Description Usage Arguments Value Author(s) Examples

Description

Calculates the volumes of stock PCR reagents required for a single template

Usage

1
pcr(stock.fwd.prim, stock.rev.prim, stock.dNTP, stock.PCR.buffer, stock.DNApol, final.fwd.prim = 0.2, final.rev.prim = 0.2, final.dNTP, final.PCR.buffer = 1, final.DNApol, stock.template, final.template, rxn.vol = 50)

Arguments

stock.fwd.prim

Stock concentration of forward primer, usually in micro moles per Litre

stock.rev.prim

Stock concentration of reverse primer, usually in micro moles per Litre

stock.dNTP

Stock concentration of dNTP mix, can be in micromoles per litre or an arbitrary concentration like 100x

stock.PCR.buffer

Stock concentration of PCR buffer

stock.DNApol

Stock concentration of DNA polymerase, can be in units per microLitre or an arbitrary concentration like 100x

stock.template

Initial concentration of template DNA, usually in ng per microlitre

final.fwd.prim

Desired concentration of forward primer in final PCR volume, defualt is 200 nM

final.rev.prim

Desired concentration of reverse primer in final PCR volume, defualt is 200 nM

final.dNTP

Desired concentration of dNTPs in the final PCR volume

final.PCR.buffer

Desired concentration of PCR Buffer in the final PCR volume, usually 1x

final.DNApol

Desired final concentration of DNA polymerase in the PCR reaction

final.template

Desired final concentration of template DNA in the PCR reaction

rxn.vol

Desired total volume of the PCR reaction, usually 50 microlitres

Value

Returns a data.frame of values corresponding to the volumes of stock solutions required.

Author(s)

Julian Spagnuolo

Examples

1
pcr(stock.fwd.prim = 6.25, stock.rev.prim = 6.25, stock.dNTP = 10, stock.PCR.buffer = 10, stock.DNApol = 50, stock.template = 20, final.fwd.prim = 0.2, final.rev.prim =0.2 , final.dNTP = 1, final.PCR.buffer = 1, final.DNApol = 1, final.template = 0.1, rxn.vol = 50)

JulianSpagnuolo/Bioscikit documentation built on May 28, 2019, 8:22 a.m.