发布时间:2019-04-22 19:57 原文链接: GeneralReference

Units

  • 1 mg = 10-3 g
    1 ug = 10-6 g
    1 ng = 10-9 g
    1 pg = 10-12 g

  • 1 kb of double stranded DNA = 660 kD 
    1 kb of single stranded DNA = 330 kD 
    1 kb of single stranded RNA = 340 kD

  • Average MW of a deoxynucleotide base = 324.5 Daltons
    Average MW of a deoxynucleotide base pair = 649 Daltons

  • 1 ug/ml of DNA = 3.08 uM phosphate
    1 ug/ml of 1 kb of DNA = 3.08 nM 5' ends
    1 mol of pBR322 = 2.83 x10+6 g.
    1 pmol of linear pBR322, 5' ends = 1.4 ug
    1 A260 unit of duplex DNA = 50 ug
    1 A260 unit of single-stranded DNA = 37 ug
    1 A260 unit of single-stranded RNA = 50 ug

  • 1 kb of DNA = 333 amino acids of coding capacity
    = 37,000 daltons

  • Densities (50% GC):

RF I (supercoiled) ds DNA1.709 g/ml
RF II (nicked) ds DNA1.54 g/ml
ss DNA1.726 g/ml
ss RNA1.90 g/ml
protein1.33 g/ml

Formulas

  • DNA melting point:

  • For duplex DNA >50 bp: 

    Tm = 81.5deg. C +16.6 log (M of NaCl) + 0.41(% GC)
    - [500/bp of shortest chain in duplex]
    - [0.65(% formamide)]

    For duplex DNA <50 bp:

    Add 2deg. C for each A or T
    Add 4deg. C for each G or C

  • Picomoles of ends:

  • pmol ends per ug linear DNA = 3030/number of bases

DNA mobility in gels

  • Migration of marker dyes in native polyacrylamide non-denaturing gels

Gel %Bromophenol blue (BP)Xylene cyanole (XC)
3.5100460
5.065260
8.045160
12.02070
20.01245
  • Migration of marker dyes in polyacrylamide denaturing gels

Gel %Bromophenol blue (BP)Xylene cyanole (XC)
5.035130
6.026106
8.01975
10.01255
20.0828
  • Relative positions of different DNA forms on Tris-acetate agarose gels

The exact distance between bands is influenced by percentage of agarose, time of electrophoresis, concentration of Ethidium bromide, degree of supercoiling and the size and complexity of the DNA.

Amino acid symbols

Alanine                         	Ala     A Arginine                        	Arg     R Asparagine                      	Asn     N Aspartic acid           		Asp     D Asparagine or Aspartic acid     	Asx     B Cysteine                        	Cys     C Glutamine                       	Gln     Q Glutamic acid                   	Glu     E Glutamine or Glutamic acid      	Glx     Z Glycine                       		Gly     G Histidine                       	His     H Isoleucine                      	Ile     I Leucine                 		Leu     L Lysine                  		Lys     K Methionine                      	Met     M Phenylalanine                   	Phe     F Proline                 		Pro     P Serine                  		Ser     S Threonine				Thr     T Tryptophan                      	Trp     W Tyrosine                        	Tyr     Y Valine                  		Val     V

Commonly used restriction enzymes and assay buffers

EnzymeIsoschizomersBufferTempRecognition SiteCloning sites
Aat II
med37GACGT/CAat II
Acc I
med37GT/(AC)(GT)ACAcc I, Cla I
Aha IIIDra Imed37TTT/AAAblunt
Alu I
med37AG/CTblunt
Asu II

37TT/CGAAAcc I, Cla I
Ava I
med37C/YCGRGSal I, Xho I, Xma I
Ava II
med37G/G(AT)CC
Bal I
low37TGG/CCAblunt
BamH1
med37G/GATCCBamH1, Bgl II
Bgl I
med37GCCN4/NGGC
Bgl II
low37A/GATCTBamH1, Bgl II
BstE II
high60G/GTNACC
BstN I
low55CC/(AT)GG
Cla I
low37AT/CGATAcc I, Cla I
Dra IAha IIIlow37TTT/AAAblunt
EcoR1
high37G/AATTCEcoR1
EcoR1*
low37/AATTEcoR1
EcoRV
med37GAT/ATCblunt
Hae I
low37(AT)GG/CC(TA)blunt
Hae II
low37RGCGC/Y
Hae III
med37GG/CCblunt
Hha ICfo I, HinP1med37GCG/CHha I
Hinc II
med37GTY/RAC blunt
Hind III
med37-55A/AGCTTHind III
Hinf I
med37G/ANTC
HinP1Cfo I, Hha Ilow37G/CGCAcc I, Cla I
Hpa I
low37GTT/AACblunt
Hpa IIMsp Ilow37C/CGGAcc I, Cla I
Kpn I
low37GGTAC/CKpn I
Mbo ISau3Amed37/GATCBamH1, Bgl II
Msp I
med37C/CGGAcc I, Cla I
Mst IFsp Ihigh37TGC/GCAblunt
Mst IIBsu36 Ihigh37CC/TNAGG
Nae I
med37GCC/CCGblunt
Nco I
high37C/CATGGNco I
Nde I
med37CA/TATGNde I
Not I
high37GC/GGCCGC
Nru I
med37TCG/CGAblunt
Pst I
med21-37CTGCA/GPst I
Pvu I
high37CGAT/CGPvu I
Pvu II
med37CAG/CTGblunt
Rsa I
med37GT/ACblunt
Sac ISst Ilow37GAGCT/CSac I, Sst I
Sal I
high37G/TCGACAva I, Sal I, Xho I
Sau3A IMbo Imed37/G*ATCBamH1, Bgl II
Sfi I